InteractiveFly: GeneBrief

Patronin: Biological Overview | References


Gene name - Patronin

Synonyms -

Cytological map position - 54C1-54C3

Function - cytoskeletal anchor protein

Keywords - microtubule minus-end-binding protein - organization of apical microtubule network to ensure size/shape homogeneity - links epithelial polarity to folding via a microtubule-based mechanical mechanism - interacts with and recruited by Shot stop which interacts with the cell cortex through its actin-binding domain - antagonist of microtubule depolymerizing kinesin Klp10A

Symbol - Patronin

FlyBase ID: FBgn0263197

Genetic map position - chr2R:17,519,024-17,535,866

NCBI classification - CAMSAP_CKK: Microtubule-binding calmodulin-regulated spectrin-associated

Cellular location - cytoplasmic



NCBI links: EntrezGene, Nucleotide, Protein
BIOLOGICAL OVERVIEW

Epithelial folding is typically driven by localized actomyosin contractility. However, it remains unclear how epithelia deform when myosin levels are low and uniform. In the Drosophila gastrula, dorsal fold formation occurs despite a lack of localized myosin changes, while the fold-initiating cells reduce cell height following basal shifts of polarity via an unknown mechanism. This study shows that cell shortening depends on an apical microtubule network organized by the CAMSAP protein Patronin. Prior to gastrulation, microtubule forces generated by the minus-end motor dynein scaffold the apical cell cortex into a dome-like shape, while the severing enzyme Katanin facilitates network remodelling to ensure tissue-wide cell size homeostasis. During fold initiation, Patronin redistributes following basal polarity shifts in the initiating cells, apparently weakening the scaffolding forces to allow dome descent. The homeostatic network that ensures size/shape homogeneity is thus repurposed for cell shortening, linking epithelial polarity to folding via a microtubule-based mechanical mechanism (Takeda, 2018).

Epithelial folding is a fundamental morphogenetic process in which two-dimensional (2D) epithelial sheets deform into 3D, folded structures. Epithelial folds form throughout animal development, giving rise to internal tissue structures and functional organ units. Folding of an epithelial monolayer can be induced by local forces generated at the site of initiation, or resulting from global stresses that buckle the tissue by exploiting intrinsic mechanical inhomogeneities. Both types of folding have been predominantly associated with modulation of myosin-dependent contractility. However, for epithelial folding events that lack overt myosin changes, it remains unclear what active mechanism underlies tissue deformation (Takeda, 2018).

Dorsal fold formation during Drosophila gastrulation has emerged as a crucial model for alternative folding mechanisms since myosin levels are low and uniform across the tissue. Following the completion of cellularization that forms the first embryonic epithelial layer, folding begins as two stripes of initiating cells straddling across the dorsal surface at stereotypical locations become shorter than their neighbours, leading to the formation of anterior and posterior dorsal folds eventually. Prior to cell shortening, Par-1, the MARK family kinase that specifies the basal-lateral membrane, retracts its apical margin and becomes downregulated, resulting in basal repositioning of adherens junctions in the initiating cells. Following such basal polarity shifts, the apices of initiating cells shrink and descend below the embryonic surface. How basal polarity shifts result in shrinkage and descent of the apices to reduce cell height was not known. As Par-1 has recently been shown to control the localization of the microtubule minus-end protector Patronin, this study investigated the possibility that Patronin-dependent modulation of the microtubule network underlies dorsal fold initiation (Takeda, 2018).

This study has identified a non-centrosomal microtubule network that is anchored and crosslinked to the apical cortex to exert pushing forces, thereby scaffolding the spherical apical shape of the epithelial cells in the Drosophila embryo prior to gastrulation. The data suggest that this microtubule network possesses an intrinsic, feedback-dependent remodelling mechanism that helps dampen the mechanical noises arising within the tissue, thus ensuring size homogeneity, echoing previous reports on microtubule-dependent cell size homeostasis. As gastrulation commences, coupling to the basal polarity shifts repurposes this homeostatic network for cell shortening (Takeda, 2018).

The microtubule forces exerted by the apical network must exist prior to, and in the opposite direction to, the movement of dome descent, thus contrasting with myosin-dependent apical constriction whose amplitude and directionality positively correlate with cell shape changes. It is proposed that dome descent in the dorsal fold system may be induced by residual stresses that compress inward and initially counterbalance the microtubule-dependent outward pushing forces, but become dominant as the basal redistribution of Patronin weakens the outward forces. This model is mechanically similar to the shape control of red blood cells where the microtubule-based marginal band pushes the cortex to counterbalance cortical tension generated by the Spectrin/actin-based membrane skeleton. It is possible that Spectrin and/or additional microtubule/actin dual linkers, such as the spectraplakin protein Shot, underlie the hypothetical counterbalancing stresses, and it is worth noting that recent work implicated links between Spectrin/Shot and Patronin/CAMSAP. Thus, in the context of the current model, Par-1 downregulation initially causes a transient expansion of the apical volume as a result of basal shifts of junctions. The expanded apical dome probably becomes mechanically imbalanced due to basal redistribution of the Patronin-anchored microtubule network, such that the pre-existing cortical compression stresses shrink the apex to drive dome descent. When such an imbalance occurs in a localized manner, a mechanical weak point could be produced to allow for initiation of folding. Conversely, global perturbation via stiffening (overexpression of Patronin) or softening (knockdown of Dynein) of the cortices probably causes a uniform deviation of such a balanced state, which instead blocks folding (Takeda, 2018).

The Patronin/CAMSAP proteins have emerged as key regulators of the microtubule minus ends, particularly for the non-centrosomal microtubules. During early cellularization prior to its transitioning to the apical cortex, however, Patronin is localized to the centrosome and appears to be required for the anchorage of centrioles and nucleus. Similar phenotypes have also been reported for the mammalian intestinal epithelia. These data could suggest an additional function on the centrosomal microtubules, which may also account for Patronin's involvement in the apical translocation of Bazooka. Intriguingly, junctional positioning in the patronin bazooka double RNAi embryos exhibits both a wide basal spread and an apical shift, respective features observed in the patronin and bazooka single knockdowns. Thus, Patronin may be required for junctional positioning in a Bazooka-independent manner. Moreover, the double RNAi embryos do not fold, even though the junctions are differentially positioned. It may be possible that the tissue-level mechanical coupling via junctions becomes defective due to the loss of Bazooka, resulting in ineffective transmission of tensile or compressive forces necessary for folding (Takeda, 2018).

The data implicate a cell shape control function for the Patronin/CAMSAP proteins in the epithelial system. The apical microtubule network that was observed resembles a similar structure previously described in the mammalian epithelial cells and could be a common feature for some epithelial systems. Highly pliable epithelial cells owing to low actomyosin-based rigidity such as those found on the dorsal side of the Drosophila gastrula may depend on the microtubule-based forces to define their shape, contrasting with epithelial contexts where cortical actomyosin forces dominate and cell shapes themselves dictate the spatial arrangement of microtubule filaments, but not vice versa. Evidence has emerged in certain contexts wherein actomyosin and microtubule networks are both involved in cell shape changes for the initiation of folding. It will be interesting to see whether context-specific interplay between these two mechanical systems underlies the formation of distinct morphological features (Takeda, 2018).

Scaling of cytoskeletal organization with cell size in Drosophila

Spatially organized macromolecular complexes are essential for cell and tissue function, but the mechanisms that organize micron-scale structures within cells are not well understood. Microtubule-based structures such as mitotic spindles scale with cell size, but less is known about the scaling of actin structures within cells. Actin-rich denticle precursors cover the ventral surface of the Drosophila embryo and larva and provide templates for cuticular structures involved in larval locomotion. Using quantitative imaging and statistical modeling, denticle number and spacing were demonstrated to scale with cell size over a wide range of cell lengths in embryos and larvae. Denticle number and spacing are reduced under space-limited conditions, and both features robustly scale over a ten-fold increase in cell length during larval growth. The relationship between cell length and denticle spacing can be recapitulated by specific mathematical equations in embryos and larvae, and accurate denticle spacing requires an intact microtubule network and the microtubule minus-end-binding protein, Patronin. These results identify a novel mechanism of microtubule-dependent actin scaling that maintains precise patterns of actin organization during tissue growth (Spencer, 2017).

Patronin/Shot cortical foci assemble the noncentrosomal microtubule array that specifies the Drosophila anterior-posterior axis

Noncentrosomal microtubules play an important role in polarizing differentiated cells, but little is known about how these microtubules are organized. This study identified the spectraplakin, Short stop (Shot), as the cortical anchor for noncentrosomal microtubule organizing centers (ncMTOCs) in the Drosophila oocyte. Shot interacts with the cortex through its actin-binding domain and recruits the microtubule minus-end-binding protein, Patronin, to form cortical ncMTOCs. Shot/Patronin foci do not co-localize with gamma-tubulin, suggesting that they do not nucleate new microtubules. Instead, they capture and stabilize existing microtubule minus ends, which then template new microtubule growth. Shot/Patronin foci are excluded from the oocyte posterior by the Par-1 polarity kinase to generate the polarized microtubule network that localizes axis determinants. Both proteins also accumulate apically in epithelial cells, where they are required for the formation of apical-basal microtubule arrays. Thus, Shot/Patronin ncMTOCs may provide a general mechanism for organizing noncentrosomal microtubules in differentiated cells (Nashchekin, 2016).

The recent discovery of the Patronin family of MT minus-end-binding proteins, consisting of Patronin in Drosophila, CAMSAP1, 2, and 3 in mammals, and PTRN-1 in worms, has begun to reveal how the minus ends of noncentrosomal MTs are organized and maintained (Akhmanova, 2015; Baines, 2009; Goodwin, 2010: Marcette, 2014; Meng, 2008; Richardson, 2014). The Patronins recognize and stabilize free MT minus ends by protecting them from depolymerization (Goodwin, 2010; Hendershott, 2014; Jiang, 2014). Patronins appear to play a particularly important role in organizing MTs in differentiated cells. CAMSAP3 localizes to the apical domain in epithelial cells, where it is required for the formation of the apical-basal array of MTs (Tanaka, 2012; Toya, 2016, Zheng, 2013). CAMSAP2 stabilizes neuronal MTs in axon and dendrites, and its knockdown leads to defects in axon specification and dendritic branch formation (Yau, 2014). Similarly, Caenorhabditis elegans PTRN-1 is required for normal neurite morphology and axon regeneration (Chuang, 2014; Marcette, 2014; Richardson, 2014). The function of Drosophila Patronin has only been examined in cultured S2 cells, where its depletion leads to a decrease in MT number and an increase in free moving MTs (Goodwin, 2010; Nashchekin, 2016 and references therein).

The polarized arrangement of the MTs in the Drosophila oocyte depends on the posterior crescent of the Par-1 kinase, which excludes MT minus ends from the posterior cortex (Doerflinger, 2010; Parton, 2011). This study shows that Par-1 acts by preventing the association of Shot with the posterior actin cortex, thereby restricting the formation of noncentrosomal MTOCs to the anterior and lateral cortex. Computer modeling has shown that this asymmetric localization of MT minus ends is sufficient to explain the formation of the weakly polarized MT network that directs the transport of oskar mRNA to the posterior pole. Thus, the regulation of the interaction of Shot with the cortex by Par-1 transmits cortical PAR polarity into the polarization of the MT cytoskeleton that localizes the axis determinants (Nashchekin, 2016).

The mechanism by which Par-1 excludes Shot is unknown. The interaction of Shot with the cortex depends on its N-terminal calponin homology domains, which bind to F-actin. Thus, Par-1 could phosphorylate Shot to inhibit its binding to the cortex. If this is the case, Par-1 would have to modify the activity or accessibility of the N-terminal ABD of Shot, as this domain recapitulates the posterior exclusion and cortical recruitment of the full-length protein. Phosphorylation of the ABD by Par-1 was not detected in vitro, however, and it seems more likely that Par-1 acts by modifying the cortex to block the binding of Shot (Nashchekin, 2016).

Shot and its vertebrate ortholog, MACF1, have previously been shown to interact with the MT plus-end tracking protein EB1 through their C-terminal SxIP motifs and with the MT lattice through their Gas2 and C-terminal domains (Nashchekin, 2016).

The current results indicate that in addition to binding to MT plus ends and to the MT lattice, Shot also interacts with MT minus ends through its association with the Patronin/Katanin complex. The exact nature of the interaction between Shot and the Patronin complex is unclear, but Shot was found to interact with Katanin 60 in a high-throughput yeast two-hybrid screen. Thus, one possibility is that Katanin acts as a link between Shot and Patronin. Since Shot is exclusively cortical in the oocyte, the protein does not appear to bind to MT plus ends or along the body of MTs in this system. It will therefore be interesting to investigate whether the different modes of MT binding by Shot are mutually exclusive and how this is regulated (Nashchekin, 2016).

Several models have been proposed to explain the formation of noncentrosomal MTs. Upon centrosome inactivation in postmitotic Drosophila tracheal cells and C. elegans intestinal cells, γ-TuRC complexes and other pericentriolar material (PCM) components are released from the centrosome and transported toward the apical membrane, where they nucleate MT. Whole MTs released from the centrosome can also be delivered and anchored to the apical domain or cell junctions by Ninein. Alternatively, new MTs can grow from MT ends generated by severing enzymes, a mechanism that is thought to be important in plant cells and neurons (Nashchekin, 2016).

This study presents evidence that this latter mechanism is responsible for the formation of the MT array that directs Drosophila axis formation. Firstly, Shot/Patronin ncMTOCs contain stable minus ends even after treatment with the MT-depolymerizing drug, colcemid, as shown by the persistent recruitment of Tau-GFP and EB1-GFP to these foci. This is consistent with the ability of Patronin and CAMPSAPs to capture and stabilize minus ends of single MTs in vitro and in cells. Secondly, MTs start to grow out in all directions from the Shot/Patronin foci immediately after colcemid inactivation. Indeed all visible growing MTs emanate from Patronin foci, indicating that they are the principal source of MTs in the oocyte. Thirdly, the foci contain no detectable γ-tubulin and do not co-localize with PCM proteins. This is consistent with observations in Caco-2 cells, which showed that CAMSAP2 and CAMSAP3 do not co-localize with γ-tubulin and in the C. elegans epidermis, where PTRN-1 and γ-tubulin function in parallel pathways to assemble circumferential MTs (Nashchekin, 2016).

Taken together, these results suggest a model in which the Shot/Patronin foci act as ncMTOCs by capturing and stabilizing MT minus-end stumps that then act as templates for new MT growth. One attractive feature of this model is that it uncouples MT organization from MT nucleation in both space and time. The Shot/Patronin complex bypasses the need to continually nucleate new MTs by preventing existing microtubules from completely depolymerizing. Thus, once a cell has nucleated sufficient MTs, it can maintain and reorganize its MT cytoskeleton by stabilizing MT minus-end stumps in appropriate locations and using these, rather than the γ-tubulin ring complex, to provide the seeds from which new MTs grow. The number of MTs can even increase in the absence of new MT nucleation if MT-severing proteins chop up existing MTs to produce new minus ends that can then be captured and stabilized. The presence of the severing protein, Katanin, in the Shot/Patronin foci is intriguing in this context, as it raises the possibility that it severs existing MTs to provide a local source of minus ends for Patronin to capture (Nashchekin, 2016).

Shot and Patronin also co-localize at the apical cortex of the epithelial follicle cells, where they are required for apical-basal MT organization. This consistent with the recent observation that CAMSAP3 is required for the recruitment of MT minus ends to the apical cortex of mammalian intestinal epithelial cells (Toya, 2016). Thus, this function of Patronin has been evolutionarily conserved. Furthermore, the similarities between roles of Shot and Patronin in the oocyte and the follicle cells suggest that the complex may provide a general mechanism for organizing noncentrosomal MTs. The relationship between Shot and Patronin is different in the follicle cells compared with the oocyte, however, as Shot is not required for the apical recruitment of Patronin. Nevertheless, loss of either protein produces a very similar loss of apical MT and a reduction in overall MT density. Although it cannot be ruled out that they act in parallel pathways, this observation suggests that they collaborate to anchor MTs to the apical cortex. The combination of Patronin binding to the MT minus ends and Shot binding to the MT lattice may therefore provide a robust anchor to retain MTs at the apical cortex (Nashchekin, 2016).

Shot and Patronin polarise microtubules to direct membrane traffic and biogenesis of microvilli in epithelia

In epithelial tissues, polarisation of microtubules and actin microvilli occurs along the apical-basal axis of each cell, yet how these cytoskeletal polarisation events are coordinated remains unclear. This study examines the hierarchy of events during cytoskeletal polarisation in Drosophila and human epithelia. Core apical-basal polarity determinants polarise the Spectrin cytoskeleton to recruit the microtubule-binding proteins Patronin (CAMSAP1/2/3 in humans) and Shortstop (Shot; MACF1/BPAG1 in humans) to the apical membrane domain. Patronin and Shot then act to polarise microtubules along the apical-basal axis to enable apical transport of Rab11 endosomes by the Nuf-Dynein microtubule motor complex. Finally, Rab11 endosomes are transferred to the MyoV actin motor to deliver the key microvillar determinant Cadherin99C to the apical membrane to organise the biogenesis of actin microvilli (Khanal, 2016).

These results reveal a mechanism linking determinants of cell polarity with stepwise polarisation of the spectrin cytoskeleton, microtubule cytoskeleton and biogenesis of actin microvilli through apical trafficking of Cad99C. The results suggest that polarisation of the apical spectrin βH-Spectrin is dependent on polarity determinants, likely through interactions with the FERM domain proteins and the apical polarity determinant Crb. The spectraplakin Shot is highly similar to βH-Spectrin, and is able to bind to and colocalise with it at the apical domain of epithelial cells, suggesting that the two proteins might have a similar function. βH-Spectrin is linked to microtubules through Patronin, whereas Shot can directly bind microtubules. Consequently, redundancy is anticipated between βH-Spectrin and Shot, or between Patronin and Shot. Accordingly, this study found that mutation of βH-spectrin only had a mild phenotype, whereas mutation of α-spectrin simultaneously disrupted both pairs of proteins in parallel and caused a drastic phenotype, completely disrupting the apical trafficking of Cad99C and microvillar biogenesis. More importantly, double mutants for shot and βH-spectrin had a more severe effect on microtubule and Cad99C localisation than either alone, therefore demonstrating that the two proteins act in a redundant fashion (Khanal, 2016).

Downstream of the spectrin cytoskeleton, Patronin and Shot are required in parallel to drive apical-basal polarisation of microtubules, which is then responsible for orienting the apical transport of Cad99C, within Rab11 endosomes, by the Dynein motor protein. Eliminating microtubules from cells by overexpressing Katanin60 results in loss of Nuf-Dynein-based apical Rab11 endosome transport and failure to efficiently deliver Cad99C to the apical membrane. The effect on Cad99C polarisation is not an indirect effect of loss of polarity due to impaired Rab11 and Dynein function in localising the apical polarity determinant Crumbs to the apical membrane because, firstly, polarity is maintained in cells expressing Rab11 or Dynein RNAi, as indicated by the normal localisation of aPKC and, secondly, loss of Crb does not strongly affect cell polarity in the follicle cell epithelium owing to redundancy with Bazooka. The results indicate that even under conditions with severe depletion of microtubules, the overall shape of the follicle cell epithelium is relatively normal, indicating that polarised microtubules are required to influence formation of apical microvilli, rather than for other functions of the actin cytoskeleton in epithelial cells. Similarly, no strong effects are seen on cell shape upon loss of either Patronin or Shot (or both), raising questions over the claimed requirement for Patronin homologs and microtubules in formation or maintenance of adherens junctions epithelial cells in culture (Khanal, 2016).

The final step in delivery of Cad99C to the apical membrane also requires actin-based transport through the action of Rip11-MyoV complex. Compromising normal MyoV function in Drosophila follicle cells by expressing a dominant-negative version of the protein, results in loss of Rab11 polarisation from the apical membrane and its abnormal accumulation in the sub-apical region. This phenotype in Drosophila shows similarities with the human microvillus inclusion disease, where mutations in the Myo5b gene also cause loss of Rab11 endosomes from the apical membrane (Khanal, 2016).

In summary, these results reveal how the spectrin cytoskeleton acts to polarise microtubules in epithelial cells, and how polarised microtubules then direct trafficking of Rab11 endosomes carrying Cad99C to the apical membrane. This process relies on a hierarchy of events, and disruption at any stage can lead to failure in delivering Cad99C to the apical membrane, resulting in defective biogenesis of microvilli. These findings are directly relevant to human diseases such as Usher's Syndrome Type 1 and microvillus inclusion disease, helping to outline the molecular and cellular basis for these conditions (Khanal, 2016).

Polarized endosome dynamics by spindle asymmetry during asymmetric cell division

During asymmetric division, fate determinants at the cell cortex segregate unequally into the two daughter cells. It has recently been shown that Sara (Smad anchor for receptor activation) signalling endosomes in the cytoplasm also segregate asymmetrically during asymmetric division (Coumailleau, 2009; Loubéry, 2014). Biased dispatch of Sara endosomes mediates asymmetric Notch/Delta signalling during the asymmetric division of sensory organ precursors in Drosophila. In flies, this has been generalized to stem cells in the gut and the central nervous system, and, in zebrafish, to neural precursors of the spinal cord. However, the mechanism of asymmetric endosome segregation is not understood. This study shows that the plus-end kinesin motor Klp98A targets Sara endosomes to the central spindle, where they move bidirectionally on an antiparallel array of microtubules. The microtubule depolymerizing kinesin Klp10A and its antagonist Patronin generate central spindle asymmetry. This asymmetric spindle, in turn, polarizes endosome motility, ultimately causing asymmetric endosome dispatch into one daughter cell. This mechanism was demonstrated by inverting the polarity of the central spindle by polar targeting of Patronin using nanobodies (single-domain antibodies). This spindle inversion targets the endosomes to the wrong cell. These data uncover the molecular and physical mechanism by which organelles localized away from the cellular cortex can be dispatched asymmetrically during asymmetric division (Derivery, 2015).

Klp98A was first identified as the kinesin mediating Sara endosome motility during sensory organ precursor (SOP) division. Klp98A is the Drosophila homologue of mammalian KIF16B, an early endosomal kinesin containing a phosphatidylinositol 3-phosphate-binding PX domain. Indeed, Klp98A localizes to Sara-positive early endosomes (Derivery, 2015).

During SOP division, Klp98A-GFP-positive Sara endosomes segregate to the pIIa daughter, but not the pII (Coumailleau, 2009; Loubéry, 2014). Sara endosomes were monitored by following Delta 20 min after internalization (iDelta20) through an improved antibody internalization assay. iDelta20 parallels Sara endosome dynamics in the controls and mutants studied here (in vivo and primary cultures. Like KIF16B, purified Klp98A binds specifically to phosphatidylinositol 3-phosphate and is a plus-end-directed motor whose velocity is 0.76 ± 0.02 μm s-1 (Derivery, 2015).

To study Klp98A function, deletions within the motor domain (Klp98AΔ6, Klp98AΔ7 and Klp98AΔ8, 6, 7 and 8-base-pair deletions, respectively) and a clean coding sequence deletion (Klp98AΔ47). Except Klp98AΔ6, all are protein nulls. In Klp98A-, Sara endosomes move diffusively. Therefore, Klp98A mediates Sara endosome motility (Derivery, 2015).

In wild-type cells, Sara endosomes move on microtubules to the Pavarotti-positive central spindle and, late in cytokinesis, to pIIa. Spindle microtubule plus-ends are oriented towards the equator, explaining central spindle endosomal targeting by a plus-end motor. Indeed, Sara endosome central spindle targeting fails in Klp98A- mutants. Importantly, in Klp98A- mutants and upon RNAi-mediated Klp98A knockdown, endosomes are symmetrically dispatched (Derivery, 2015).

Klp98A-mediated motility contributes to cell fate assignation through asymmetric Notch signalling, but this activity is redundantly covered by Neuralized and Numb (Fürthauer, 2009). Indeed, Klp98A-;pnr > neurRNAidouble mutants show a synergistic fate assignation phenotype: the notum is largely void of bristles. Conversely, Klp98A;Numb double mutants strongly suppress the diagnostic Numb- multiple socket phenotype. Therefore, having established the role of Klp98A motility in Notch signalling, this study focused on the mechanisms orchestrating asymmetric motility (Derivery, 2015).

Central spindle targeting of Sara endosomes precedes asymmetric segregation to pIIa. Focus was therefore placed on Sara endosome motility with respect to the central spindle reference frame. The central spindle is composed of the Pavarotti-positive core (containing antiparallel microtubules) plus the microtubules emanating from it. The Pavarotti core was automatically tracked, defining a 2D cartesian reference frame whose origin is the Pavarotti centroid and whose x axis is the pIIb-pIIa axis. This also defines a Pavarotti width (PW) and length (PL, the length of the microtubule antiparallel array (Derivery, 2015).

Sara endosomes were tracked with respect to this reference frame (with 160 nm accuracy. Automatic tracking and spatio-temporal registration provided a large data set (2,897 traces) from which a spatio-temporal density plot of endosomes at the central spindle was generated. For 500 s, endosomes remain mostly within the Pavarotti region. Remarkably, at the central spindle, motility along the x axis is bidirectional. Motility along the y axis merely follows PW contraction, consistent with motility along central spindle microtubules, parallel to the x axis. Velocities are similar towards pIIa and pIIb and slower than in vitro, possibly due to crowding by microtubule-associated proteins (Derivery, 2015).

Confinement within the Pavarotti region and bidirectional movement are both consistent with a plus-end motor switching direction on antiparallel microtubules. On single microtubules, Klp98A-bound quantum dots always maintain their directionality when resuming after a pause. It was asked whether Klp98A could switch direction in an antiparallel bundle. In an in vitro reconstitution assay, Klp98A-bound quantum dots move bidirectionally within antiparallel MAP65-1-mediated microtubule arrays. 68% tracks change direction after pausing. Therefore, Klp98A supports bidirectional motility in antiparallel array (Derivery, 2015).

Notably, in vivo, bidirectional endosome motility is asymmetric: the residence time in pIIa is 1.8-fold longer than in pIIb. Consistently, the spatio-temporal density plot is asymmetric. Furthermore, tracks overshoot beyond the Pavarotti region more frequently into pIIa (Derivery, 2015).

Eventually, endosomes depart from central spindle microtubules into the cytoplasm and therefore move also on the y axis. The longer pIIa residence time and higher pIIa overshoot frequency make this final departure asymmetric, explaining the biased segregation into pIIa. Therefore, asymmetric endosome motility at the central spindle underlies asymmetric dispatch to pIIa (Derivery, 2015).

It was then asked whether the central spindle itself is asymmetric. Using Pavarotti spatio-temporal registration, an 'average cell' was generated to map the densities of the microtubule markers Jupiter and SiR-tubulin (microtubule markers), Patronin (Goodwin, 2010) (minus-end), and Pavarotti (plus-ends/antiparallel overlap). This 'average cell' reveals a polarity map of the central spindle consistent with electron microscopy reports: plus-ends are in the middle and minus-ends on the outer side. Microtubule densities in general, and Patronin in particular, are ~20% higher on the pIIb side. This asymmetry depends on Par complex activity, and is absent in neighbouring cells dividing symmetrically, but this seems independent of central spindle or endosomal asymmetry (Derivery, 2015).

Microtubule asymmetry builds up during anaphaseB, concomitant with biased endosome motility, while, earlier, the metaphase spindle is symmetrical. During anaphaseB, the central spindle shrinks by microtubule depolymerization through depolymerizing kinesins like Klp10A, among other factors. Depolymerization dynamics are asymmetric: microtubule loss is faster in pIIa. This could be explained by Patronin enrichment in the central spindle pIIb outer side where it binds to minus-ends, counteracting Klp10A-mediated depolymerization (Goodwin, 2010; Wang, 2013; Hendershott, 2014; Jiang, 2014; Derivery, 2015 and references therein).

Indeed, Klp10A/Patronin control asymmetric microtubule depolymerization: their depletion abolishes spindle asymmetry. In Patronin-knockdown cells, both sides exhibit low microtubule densities characteristic of pIIa, consistent with Patronin pIIb enrichment in wild type and its activity against depolymerization. Conversely, upon knockdown of Klp10A, both sides exhibit high microtubule densities resembling pIIb (Derivery, 2015).

The parallelism between central spindle asymmetry and asymmetric endosome motility suggests that spindle asymmetry causes biased motility. Indeed, endosome motility at the central spindle and, therefore, segregation become symmetric in Klp10A- and Patronin-knockdowns, while early central spindle targeting is normal. This uncovers a quantitative correlation between spindle and endosomal asymmetry (Derivery, 2015).

Together, a plus-end motor and microtubule plus-ends facing the centre explain why a higher pIIb microtubule density (~20% enrichment) targets endosomes to pIIa (~80% pIIa, that is, >300% enrichment). In other words, endosomes move away from higher microtubule densities in pIIb (Derivery, 2015).

Based on a theoretical model of plus-end endosomal motility on an antiparallel, asymmetric microtubule overlap, the steady-state endosome distribution is

Where PpIIa, PpIIb, the probabilities for an endosome to be on either side of the antiparallel overlap; ρa, ρb, microtubule densities in pIIa/pIIb, respectively; kon, koff, microtubule association/dissociation constants of the motor, respectively; v, the endosome motor-driven velocity; D, the diffusion coefficient of endosomes detached from microtubules; and l, the antiparallel overlap length (Derivery, 2015).

To generate this inverted spindle, 'nanobody assay' was established based on GFP-binding-peptide (GBP)-Pon, a nanobody fused to the Pon localization domain. GBP-Pon traps GFP-Patronin away from the spindle at the pIIb cortex thereby reducing, specifically in pIIb, Patronin-dependent protection against central spindle depolymerization. This inverts spindle asymmetry, which consequently inverts endosomal asymmetry. SiR-Tubulin and endogenous acetyl-tubulin stainings confirmed this spindle inversion (Derivery, 2015).

Interestingly, this assay generates a phenotypic series of different levels of spindle reversal and their corresponding endosomal reversals. These data fall on the theoretical curve obtained with independently measured parameters. Therefore these results uncover the quantitative dependence of asymmetric endosome targeting on spindle asymmetry (Derivery, 2015).

This study has identified Klp10A/Patronin as the machinery generating spindle asymmetry, which is read out by Klp98A to achieve asymmetric targeting of signalling endosomes. Asymmetric endosomal targeting contributes in turn to asymmetric cell fate assignation, confirming previous reports in flies and fish. These data thus uncover a mechanism by which intracellular cargoes in general, and signalling endosomes in particular, can be targeted to one of the daughter cells during asymmetric division (Derivery, 2015).

How could then other cargoes segregate symmetrically, if the spindle is asymmetric? Asymmetric targeting would only be efficient if kon, koff and v are optimized to amplify the mild asymmetry of the spindle, otherwise concealed by noise sources in the cell. More generally, plus- and minus-end motors are present simultaneously in the same vesicle and thereby may counteract each other to achieve symmetrical dispatch (a sort of 'tug of war'). Therefore, the precise landscape of microtubule polarity trails combined with the right cocktail of motors in vesicles provides the plasticity required to generate the plethora of molecular spatial patterns observed in polarized cells (Derivery, 2015).

Regulation of microtubule minus-end dynamics by CAMSAPs and Patronin

The microtubule (MT) cytoskeleton plays an essential role in mitosis, intracellular transport, cell shape, and cell migration. The assembly and disassembly of MTs, which can occur through the addition or loss of subunits at the plus- or minus-ends of the polymer, is essential for MTs to carry out their biological functions. A variety of proteins act on MT ends to regulate their dynamics, including a recently described family of MT minus-end binding proteins called calmodulin-regulated spectrin-associated protein (CAMSAP)/Patronin/Nezha. Patronin, the single member of this family in Drosophila, was previously shown to stabilize MT minus-ends against depolymerization in vitro and in vivo. This study shows that all three mammalian CAMSAP family members also bind specifically to MT minus-ends and protect them against kinesin-13-induced depolymerization. However, these proteins differ in their abilities to suppress tubulin addition at minus-ends and to dissociate from MTs. CAMSAP1 does not interfere with polymerization and tracks along growing minus-ends. CAMSAP2 and CAMSAP3 decrease the rate of tubulin incorporation and remain bound, thereby creating stretches of decorated MT minus-ends. By using truncation analysis, this study found that somewhat different minimal domains of CAMSAP and Patronin are involved in minus-end localization. However, in both cases, a highly conserved C-terminal domain and a more variable central domain were found to cooperate to suppress minus-end dynamics in vitro, and both regions are required to stabilize minus-ends in Drosophila S2 cells. These results show that members of the CAMSAP/Patronin family all localize to and protect minus-ends but have evolved distinct effects on MT dynamics (Hendershott, 2014).

Patronin regulates the microtubule network by protecting microtubule minus ends

Tubulin assembles into microtubule polymers that have distinct plus and minus ends. Most microtubule plus ends in living cells are dynamic; the transitions between growth and shrinkage are regulated by assembly-promoting and destabilizing proteins. In contrast, minus ends are generally not dynamic, suggesting their stabilization by some unknown protein. This study has identified Patronin (also known as ssp4) as a protein that stabilizes microtubule minus ends in Drosophila S2 cells. In the absence of Patronin, minus ends lose subunits through the actions of the Kinesin-13 microtubule depolymerase, leading to a sparse interphase microtubule array and short, disorganized mitotic spindles. In vitro, the selective binding of purified Patronin to microtubule minus ends is sufficient to protect them against Kinesin-13-induced depolymerization. It is proposed that Patronin caps and stabilizes microtubule minus ends, an activity that serves a critical role in the organization of the microtubule cytoskeleton (Goodwin, 2010).


Functions of Patronin orthologs in other species

CAMSAP3 orients the apical-to-basal polarity of microtubule arrays in epithelial cells

Polarized epithelial cells exhibit a characteristic array of microtubules that are oriented along the apicobasal axis of the cells. The minus-ends of these microtubules face apically, and the plus-ends face toward the basal side. The mechanisms underlying this epithelial-specific microtubule assembly remain unresolved, however. Using mouse intestinal cells and human Caco-2 cells, this study shows that the microtubule minus-end binding protein CAMSAP3 (calmodulin-regulated-spectrin-associated protein 3) plays a pivotal role in orienting the apical-to-basal polarity of microtubules in epithelial cells. In these cells, CAMSAP3 accumulated at the apical cortices, and tethered the longitudinal microtubules to these sites. Camsap3 mutation or depletion resulted in a random orientation of these microtubules; concomitantly, the stereotypic positioning of the nucleus and Golgi apparatus was perturbed. In contrast, the integrity of the plasma membrane was hardly affected, although its structural stability was decreased. Further analysis revealed that the CC1 domain of CAMSAP3 is crucial for its apical localization, and that forced mislocalization of CAMSAP3 disturbs the epithelial architecture. These findings demonstrate that apically localized CAMSAP3 determines the proper orientation of microtubules, and in turn that of organelles, in mature mammalian epithelial cells (Toya, 2016).

DAPK interacts with Patronin and the microtubule cytoskeleton in epidermal development and wound repair

Epidermal barrier epithelia form a first line of defense against the environment, protecting animals against infection and repairing physical damage. In C. elegans, death-associated protein kinase (DAPK-1) (see Drosophila Drak, which promotes phosphorylation of Spaghetti squash at sites known to stimulate actomyosin contractility) regulates epidermal morphogenesis, innate immunity and wound repair. This study found that DAPK-1 maintains epidermal tissue integrity through regulation of the microtubule (MT) cytoskeleton. dapk-1 epidermal phenotypes are suppressed by treatment with microtubule-destabilizing drugs and mimicked or enhanced by microtubule-stabilizing drugs. Loss of function in ptrn-1 (see Drosophila Patronin), the C. elegans member of the Patronin/Nezha/CAMSAP family of MT minus-end binding proteins, suppresses dapk-1 epidermal and innate immunity phenotypes. Over-expression of the MT-binding CKK domain of PTRN-1 triggers epidermal and immunity defects resembling those of dapk-1 mutants, and PTRN-1 localization is regulated by DAPK-1. DAPK-1 and PTRN-1 physically interact in co-immunoprecipitation experiments, and DAPK-1 itself undergoes MT-dependent transport. These data uncover an unexpected interdependence of DAPK-1 and the microtubule cytoskeleton in maintenance of epidermal integrity (Chuang, 2016).

The microtubule minus-end-binding protein patronin/PTRN-1 is required for axon regeneration in C. elegans

Precise regulation of microtubule (MT) dynamics is increasingly recognized as a critical determinant of axon regeneration. In contrast to developing neurons, mature axons exhibit noncentrosomal microtubule nucleation. The factors regulating noncentrosomal MT architecture in axon regeneration remain poorly understood. This study reports that PTRN-1, the C. elegans member of the Patronin/Nezha/calmodulin- and spectrin-associated protein (CAMSAP) family of microtubule minus-end-binding proteins, is critical for efficient axon regeneration in vivo. ptrn-1-null mutants display generally normal developmental axon outgrowth but significantly impaired regenerative regrowth after laser axotomy. Unexpectedly, mature axons in ptrn-1 mutants display elevated numbers of dynamic axonal MTs before and after injury, suggesting that PTRN-1 inhibits MT dynamics. The CKK domain of PTRN-1 is necessary and sufficient for its functions in axon regeneration and MT dynamics and appears to stabilize MTs independent of minus-end localization. Whereas in developing neurons, PTRN-1 inhibits activity of the DLK-1 mitogen-activated protein kinase (MAPK) cascade, this study finds that, in regeneration, PTRN-1 and DLK-1 function together to promote axonal regrowth (Chuang, 2014).

Microtubule minus-end stabilization by polymerization-driven CAMSAP deposition

Microtubules are cytoskeletal polymers with two structurally and functionally distinct ends, the plus- and the minus-end. This study focused on the mechanisms underlying the regulation of microtubule minus-ends by the CAMSAP/Nezha/Patronin protein family. CAMSAP2 is shown to be required for the proper organization and stabilization of interphase microtubules and directional cell migration. By combining live-cell imaging and in vitro reconstitution of microtubule assembly from purified components with laser microsurgery, it was demonstrate that CAMSAPs regulate microtubule minus-end growth and are specifically deposited on the lattice formed by microtubule minus-end polymerization. This process leads to the formation of CAMSAP-decorated microtubule stretches, which are stabilized from both ends and serve as sites of noncentrosomal microtubule outgrowth. The length of the stretches is regulated by the microtubule-severing protein katanin, which interacts with CAMSAPs. These data thus indicate that microtubule minus-end assembly drives the stabilization of noncentrosomal microtubules and that katanin regulates this process (Jiang, 2014).

Patronin mediates a switch from kinesin-13-dependent poleward flux to anaphase B spindle elongation

Anaphase B spindle elongation contributes to chromosome segregation during Drosophila melanogaster embryo mitosis. It is proposed that this process is driven by a kinesin-5-generated interpolar microtubule (MT; ipMT) sliding filament mechanism that engages when poleward flux is turned off. This paper presents evidence that anaphase B is induced by the minus end-stabilizing protein patronin, which antagonizes the kinesin-13 depolymerase KLP10A at spindle poles, thereby switching off the depolymerization of the minus ends of outwardly sliding ipMTs to suppress flux. Although intact cortices, kinetochore MTs, and midzone augmentation are dispensable, this patronin-based change in ipMT minus-end dynamics is sufficient to induce the elongation of spindles capable of separating chromosomes (Wang, 2013).

PTRN-1, a microtubule minus end-binding CAMSAP homolog, promotes microtubule function in Caenorhabditis elegans neurons

In neuronal processes, microtubules (MTs) provide structural support and serve as tracks for molecular motors. While it is known that neuronal MTs are more stable than MTs in non-neuronal cells, the molecular mechanisms underlying this stability are not fully understood. This study used live fluorescence microscopy to show that the C. elegans CAMSAP protein PTRN-1 localizes to puncta along neuronal processes, stabilizes MT foci, and promotes MT polymerization in neurites. Electron microscopy revealed that ptrn-1 null mutants have fewer MTs and abnormal MT organization in the PLM neuron. Animals grown with a MT depolymerizing drug caused synthetic defects in neurite branching in the absence of ptrn-1 function, indicating that PTRN-1 promotes MT stability. Further, ptrn-1 null mutants exhibited aberrant neurite morphology and synaptic vesicle localization that is partially dependent on dlk-1. These results suggest that PTRN-1 represents an important mechanism for promoting MT stability in neurons (Richardson, 2014).

Microtubule minus-end binding protein CAMSAP2 controls axon specification and dendrite development

In neurons, most microtubules are not associated with a central microtubule-organizing center (MTOC), and therefore, both the minus and plus-ends of these non-centrosomal microtubules are found throughout the cell. Microtubule plus-ends are well established as dynamic regulatory sites in numerous processes, but the role of microtubule minus-ends has remained poorly understood. Using live-cell imaging, high-resolution microscopy, and laser-based microsurgery techniques, this study shows that the CAMSAP/Nezha/Patronin family protein CAMSAP2 specifically localizes to non-centrosomal microtubule minus-ends and is required for proper microtubule organization in neurons. CAMSAP2 stabilizes non-centrosomal microtubules and is required for neuronal polarity, axon specification, and dendritic branch formation in vitro and in vivo. Furthermore, non-centrosomal microtubules in dendrites were found to be largely generated by gamma-Tubulin-dependent nucleation. A two-step model is proposed in which gamma-Tubulin initiates the formation of non-centrosomal microtubules and CAMSAP2 stabilizes the free microtubule minus-ends in order to control neuronal polarity and development (Yau, 2014).

Microtubule minus-end stabilization by polymerization-driven CAMSAP deposition

Microtubules are cytoskeletal polymers with two structurally and functionally distinct ends, the plus- and the minus-end. This study focused on the mechanisms underlying the regulation of microtubule minus-ends by the CAMSAP/Nezha/Patronin protein family. CAMSAP2 was shown to be required for the proper organization and stabilization of interphase microtubules and directional cell migration. By combining live-cell imaging and in vitro reconstitution of microtubule assembly from purified components with laser microsurgery, it was demonstrated that CAMSAPs regulate microtubule minus-end growth and are specifically deposited on the lattice formed by microtubule minus-end polymerization. This process leads to the formation of CAMSAP-decorated microtubule stretches, which are stabilized from both ends and serve as sites of noncentrosomal microtubule outgrowth. The length of the stretches is regulated by the microtubule-severing protein katanin, which interacts with CAMSAPs. These data thus indicate that microtubule minus-end assembly drives the stabilization of noncentrosomal microtubules and that katanin regulates this process (Jiang, 2014).

The Caenorhabditis elegans microtubule minus-end binding homolog PTRN-1 stabilizes synapses and neurites

Microtubule dynamics facilitate neurite growth and establish morphology, but the role of minus-end binding proteins in these processes is largely unexplored. CAMSAP homologs associate with microtubule minus-ends, and are important for the stability of epithelial cell adhesions. This study reports morphological defects in neurons and neuromuscular defects in mutants of the C. elegans CAMSAP, ptrn-1. Mechanosensory neurons initially extend wild-type neurites, and subsequently remodel by overextending neurites and retracting synaptic branches and presynaptic varicosities. This neuronal remodeling can be activated by mutations known to alter microtubules, and depends on a functioning DLK-1 MAP kinase pathway. PTRN-1 was found to localize to both neurites and synapses, and the results suggest that alterations of microtubule structures caused by loss of PTRN-1 function activates a remodeling program leading to changes in neurite morphology. A model is proposed whereby minus-end microtubule stabilization mediated by a functional PTRN-1 is necessary for morphological maintenance of neurons (Marcette, 2014).


REFERENCES

Search PubMed for articles about Drosophila Patronin

Akhmanova, A. and Steinmetz, M. O. (2015). Control of microtubule organization and dynamics: two ends in the limelight. Nat Rev Mol Cell Biol 16(12): 711-726. PubMed ID: 26562752

Baines, A. J., Bignone, P. A., King, M. D., Maggs, A. M., Bennett, P. M., Pinder, J. C. and Phillips, G. W. (2009). The CKK domain (DUF1781) binds microtubules and defines the CAMSAP/ssp4 family of animal proteins. Mol Biol Evol 26(9): 2005-2014. PubMed ID: 19508979

Chuang, M., Goncharov, A., Wang, S., Oegema, K., Jin, Y. and Chisholm, A. D. (2014). The microtubule minus-end-binding protein patronin/PTRN-1 is required for axon regeneration in C. elegans. Cell Rep 9(3): 874-883. PubMed ID: 25437544

Chuang, M., Hsiao, T.I., Tong, A., Xu, S. and Chisholm, A.D. (2016). DAPK interacts with Patronin and the microtubule cytoskeleton in epidermal development and wound repair. Elife 5. pii: e15833. PubMed ID: 27661253

Coumailleau, F., Furthauer, M., Knoblich, J. A. and Gonzalez-Gaitan, M. (2009). Directional Delta and Notch trafficking in Sara endosomes during asymmetric cell division. Nature 458(7241): 1051-1055. PubMed ID: 19295516

Derivery, E., Seum, C., Daeden, A., Loubery, S., Holtzer, L., Julicher, F. and Gonzalez-Gaitan, M. (2015). Polarized endosome dynamics by spindle asymmetry during asymmetric cell division. Nature 528: 280-285. PubMed ID: 26659188

Doerflinger, H., Vogt, N., Torres, I. L., Mirouse, V., Koch, I., Nusslein-Volhard, C. and St Johnston, D. (2010). Bazooka is required for polarisation of the Drosophila anterior-posterior axis. Development 137(10): 1765-1773. PubMed ID: 20430751

Furthauer, M. and Gonzalez-Gaitan, M. (2009). Endocytic regulation of notch signalling during development. Traffic 10(7): 792-802. PubMed ID: 19416471

Goodwin, S. S. and Vale, R. D. (2010). Patronin regulates the microtubule network by protecting microtubule minus ends. Cell 143(2): 263-274. PubMed ID: 20946984

Hendershott, M. C. and Vale, R. D. (2014). Regulation of microtubule minus-end dynamics by CAMSAPs and Patronin. Proc Natl Acad Sci U S A 111(16): 5860-5865. PubMed ID: 24706919

Jiang, K., Hua, S., Mohan, R., Grigoriev, I., Yau, K. W., Liu, Q., Katrukha, E. A., Altelaar, A. F., Heck, A. J., Hoogenraad, C. C. and Akhmanova, A. (2014). Microtubule minus-end stabilization by polymerization-driven CAMSAP deposition. Dev Cell 28(3): 295-309. PubMed ID: 24486153

Khanal, I., Elbediwy, A., Diaz de la Loza, M.D., Fletcher, G.C. and Thompson, B.J. (2016). Shot and Patronin polarise microtubules to direct membrane traffic and biogenesis of microvilli in epithelia. J Cell Sci 129(13):2651-9. PubMed ID: 27231092

Loubéry, S., Seum, C., Moraleda, A., Daeden, A., Furthauer, M. and Gonzalez-Gaitan, M. (2014). Uninflatable and Notch control the targeting of Sara endosomes during asymmetric division. Curr Biol 24(18): 2142-2148. PubMed ID: 25155514

Marcette, J. D., Chen, J. J. and Nonet, M. L. (2014). The Caenorhabditis elegans microtubule minus-end binding homolog PTRN-1 stabilizes synapses and neurites. Elife 3: e01637. PubMed ID: 24569480

Meng, W., Mushika, Y., Ichii, T. and Takeichi, M. (2008). Anchorage of microtubule minus ends to adherens junctions regulates epithelial cell-cell contacts. Cell 135(5): 948-959. PubMed ID: 19041755

Nashchekin, D., Fernandes, A. R. and St Johnston, D. (2016). Patronin/Shot cortical foci assemble the noncentrosomal microtubule array that specifies the Drosophila anterior-posterior axis. Dev Cell 38: 61-72. PubMed ID: 27404359

Parton, R. M., Hamilton, R. S., Ball, G., Yang, L., Cullen, C. F., Lu, W., Ohkura, H. and Davis, I. (2011). A PAR-1-dependent orientation gradient of dynamic microtubules directs posterior cargo transport in the Drosophila oocyte. J Cell Biol 194(1): 121-135. PubMed ID: 21746854

Richardson, C. E., Spilker, K. A., Cueva, J. G., Perrino, J., Goodman, M. B. and Shen, K. (2014). PTRN-1, a microtubule minus end-binding CAMSAP homolog, promotes microtubule function in Caenorhabditis elegans neurons. Elife 3: e01498. PubMed ID: 24569477

Spencer, A. K., Schaumberg, A. J. and Zallen, J. A. (2017). Scaling of cytoskeletal organization with cell size in Drosophila. Mol Biol Cell 28(11):1519-1529. PubMed ID: 28404752

Tanaka, N., Meng, W., Nagae, S. and Takeichi, M. (2012). Nezha/CAMSAP3 and CAMSAP2 cooperate in epithelial-specific organization of noncentrosomal microtubules. Proc Natl Acad Sci U S A 109(49): 20029-20034. PubMed ID: 23169647

Takeda, M., Sami, M. M. and Wang, Y. C. (2018). A homeostatic apical microtubule network shortens cells for epithelial folding via a basal polarity shift. Nat Cell Biol 20(1): 36-45. PubMed ID: 29203884

Toya, M., Kobayashi, S., Kawasaki, M., Shioi, G., Kaneko, M., Ishiuchi, T., Misaki, K., Meng, W. and Takeichi, M. (2016). CAMSAP3 orients the apical-to-basal polarity of microtubule arrays in epithelial cells. Proc Natl Acad Sci U S A 113(2): 332-337. PubMed ID: 26715742

Wang, H., Brust-Mascher, I., Civelekoglu-Scholey, G. and Scholey, J. M. (2013). Patronin mediates a switch from kinesin-13-dependent poleward flux to anaphase B spindle elongation. J Cell Biol 203(1): 35-46. PubMed ID: 24100293

Yau, K. W., van Beuningen, S. F., Cunha-Ferreira, I., Cloin, B. M., van Battum, E. Y., Will, L., Schatzle, P., Tas, R. P., van Krugten, J., Katrukha, E. A., Jiang, K., Wulf, P. S., Mikhaylova, M., Harterink, M., Pasterkamp, R. J., Akhmanova, A., Kapitein, L. C. and Hoogenraad, C. C. (2014). Microtubule minus-end binding protein CAMSAP2 controls axon specification and dendrite development. Neuron 82(5): 1058-1073. PubMed ID: 24908486

Zheng, J., Furness, D., Duan, C., Miller, K. K., Edge, R. M., Chen, J., Homma, K., Hackney, C. M., Dallos, P. and Cheatham, M. A. (2013). Marshalin, a microtubule minus-end binding protein, regulates cytoskeletal structure in the organ of Corti. Biol Open 2(11): 1192-1202. PubMed ID: 24244856


Biological Overview

date revised: 24 September 2018

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