bride of sevenless: Biological Overview | Regulation | Developmental Biology | Effects of Mutation | References

Gene name - bride of sevenless

Synonyms -

Cytological map position - 96F5-14

Function - ligand of sevenless

Keywords - boss-sevenless pathway

Symbol - boss

FlyBase ID:FBgn0000206

Genetic map position - 3-90.5

Classification - seven transmembrane protein

Cellular location - cell surface



NCBI links: Precomputed BLAST | Entrez Gene |
BIOLOGICAL OVERVIEW

There are eight photoreceptor cells (R-cells) in each ommatidium (facet) of the Drosophila compound eye. The first cell to develop is the R8 cell, followed by R1 through R6, with R7 following these. The determination of R7 photoreceptor fate depends on the expression of boss in the neighboring R8 cell. What is the origin of this observation? How can the identity and genotype of each cell be determined in the developing retina?

Mosaic analysis has been used to investigate the cellular requirements for R7 development. The chaoptic (chp) mutation which, as a homozygote mutant, gives an altered cell morphology was linked to the boss mutation. Flies heterozygous for the linked mutations boss and chp were then subject to X-rays to induced mitotic recombination. Mitotic recombination gives some cells of altered morphology identifiable as boss negative cells (on the basis of their altered morphology induced by the recessive chp mutation.

About one in a hundred flies exhibits mosaic retinas, that is, some cells inherit wild type chromosomes and other cells inherit recombinante chromosomes homozygous for chp and boss mutations. The double mutant cells exist as clones in the developing retina; the cells of particular interest are found at the border of mutant and normal clones, in ommatidia that possess cells of mixed ancestory. R7 development is independent of boss expression in R1 through R6 cells in mixed ommatidia but the genotype of R8 cells has to be wild type for bossif R7 cells are to develop. Since wild type R8 could not rescue an R7 in adjacent ommatidia but only within an ommatidium, it is concluded that BOSS can act non-cell autonomously as a ligand to induce the R7 fate only over a short distance (Reinke, 1988).

The target of BOSS is the receptor Sevenless. Studies reveal that interaction of cells bearing the BOSS ligand can aggregate with cells bearing the Sevenless receptor. There is considerable interest in the fate of receptor and ligand when ligand induces receptor signaling. In the case of the BOSS-Sevenless interaction, BOSS is internalized by sev expressing cells. BOSS immunoreactivity is not found on the surface of Sevenless bearing cells in the developing retina, but rather in vesicles inside the cell. This vesicular BOSS depends on endocytosis. When endocytosis is prevented in shabire mutants (shabire codes for the Drosophila homolog of vertebrate Dynamin, involved in endocytosis), BOSS protein is not apparent in R7 cells (Krämer, 1993).

Binding of boss-expressing cells to sev-expressing cells results in a rapid increase in phosphorylation of the Sevenless receptor tyrosine kinase. A BOSS amino-terminal peptide containing only the large extracellular domain was found to act as an inhibitor of BOSS-Sevenless directed cell adhesion, preventing SEV from undergoing phosphorylation (Hart, 1993). These experiments unequivocally show that Boss, as a ligand, binds and activates SEV (Yamamoto, 1994).

BOSS structure is that of a seven pass transmembrane G protein coupled receptor. Does BOSS actually function as a G protein coupled receptor, and if so, what is its function? Could BOSS be signaling back to the cell bearing it that an R7 fate has been determined in an adjacent cell? There will be other instances of G protein coupled receptors behaving as ligands during development.

Evidence for signaling from R7 to R8 photoreceptors (reverse information flow) comes from a study of rhodopsin expression in the two cell populations. The outer (R1-R6) and inner (R7, R8) photoreceptors represent two physiologically distinct systems with two different projection targets in the brain. All cells of the primary system, R1-R6, express the same rhodopsin and are functionally identical. In contrast, the R7 and R8 photoreceptors are different from each other. They occupy anatomically precise positions, with R7 on top of R8. In fact, there are several classes of R7/R8 pairs, which differ morphologically and functionally and are characterized by the expression of one of two R7-specific opsins: rh3 or rh4. A new opsin gene has been identified, rhodopsin 5, expressed in one subclass of R8 cells. Interestingly, this subclass represents R8 cells that are directly underneath the R7 photoreceptors expressing rh3, but are never under those expressing rh4. These results confirm the existence of two subpopulations of R7 and R8 cells, which coordinate the expression of their respective rh genes. Thus, developmental signaling pathways between R7 and R8 lead to the exclusive expression of a single rhodopsin gene per cell and to the coordinate expression of another one in the neighboring cell. Consistent with this, rh5 expression in R8 disappears when R7 cells are absent (in sevenless mutant) (Papatsenko, 1997).

The most striking observation reported here is that rh5 is expressed in a subset of R8 cells, which corresponds exactly to the R7 cells expressing rh3. This makes it very likely that there is a mechanism for synchronization of rh gene expression between the two cells. Two models are suggested:

Although the absence of rh5 in sevenless mutants does not provide evidence in favor of either model, it may reflect the existence of a positive feedback loop in the interaction between the R8 and R7 cells. Since the R8 cells do not develop properly (rh5 is not expressed) in the absence of R7, a signal from R7 must be necessary to complete retinal development and for the formation of the different subclasses of ommatidia. The sevenless-boss pathway, or some part of it, may be involved in this interaction. Alternatively, a totally distinct pathway may lead to coordination of rh gene expression in R8. It is interesting to mention that physiological data indicate that there are still functional R8 cells in sevenless mutants. Indeed, a photopigment could be detected in the retina of flies lacking both functional R1-6 cells (ninaE mutants) and R7 cells (sev mutants). Thus, an unknown rhodopsin (rh6?), which is predicted to be expressed in the remaining subset of R8 cells (R8y; in concert with rh4 in R7y), could be expressed in all R8 cells in sevenless flies. If this is true, the R7 cell controls the expression of rh5 in R8, and thus the determination of the particular subclass of ommatidia. In other words, the decision for transcriptional exclusion between rh3/5 and rh4/6 would occur in R7, downstream of the boss-sev signaling and upstream of the suggested feedback control from R7 to R8. It must be noted that expression of rh3 in R8 cells of the dorsal margin is not affected in sevenless mutants. This indicates that at least some of the R8 development is not affected by R7 cells. In conclusion, the discrimination between rh3 and rh4 (and between rh5 and putative rh6) expression, and the coordination between pairs of rh genes in neighboring photoreceptors provides a powerful paradigm for studying not only the mechanism of transcriptional exclusion often found in sensory systems, but also for studying the concerted evolution of rhodopsin genes in achieving specific functions, such as color vision (Papatsenko, 1997).

Boss functions as a glucose-responding receptor: loss of boss causes abnormal energy metabolism

Glucose, one of the most important nutrients for animals, acts as a regulatory signal that controls the secretion of hormones, such as insulin, by endocrine tissues. However, how organisms respond to extracellular glucose and how glucose controls nutrient homeostasis remain unknown. This study shows that a putative Drosophila G protein-coupled receptor, previously identified as Bride of sevenless (BOSS), responds to extracellular glucose and regulates sugar and lipid metabolism. BOSS was expressed in the fat body, a nutrient-sensing tissue equivalent to mammalian liver and adipose tissues, and in photoreceptor cells. Boss null mutants had small bodies, exhibited abnormal sugar and lipid metabolism (elevated circulating sugar and lipid levels, impaired lipid mobilization to oenocytes), and were sensitive to nutrient deprivation stress. These phenotypes are reminiscent of flies defective in insulin signaling. Consistent with these findings are the observations that boss mutants have reduced PI3K activity and phospho-AKT levels, indicating that BOSS is required for proper insulin signaling. Because human G protein-coupled receptor 5B and the seven-transmembrane domain of BOSS share the same sequence, these results also have important implications for glucose metabolism in humans. Thus, this study provides insight not only into the basic mechanisms of metabolic regulation but also into the pathobiological basis for diabetes and obesity (Kohyama-Koganeya, 2008).

The discovery of an additional function of BOSS as a glucose-responding GPCR is an example in multicellular organisms of a glucose-responding GPCR that regulates glucose and lipid metabolism. Sequence similarity between the BOSS 7TM-C-terminal region and vertebrate GPRC5Bs strongly suggests that the conserved region of BOSS functions as a glucose-responding receptor. Supporting this suggestion is the observation that both BOSS and GPRC5B expressed in the larval fat body were internalized in response to glucose stimulation. The same response to glucose stimulation was also observed in GPRC5B-expressing HEK293 cells. This type of glucose-dependent internalization was specific to GPRC5B: Internalization of GPRC5D, which belongs to the same subfamily as GPRC5B, was not detected. Although evidence is provided that BOSS responds to glucose through glucose-dependent receptor-internalization, luciferase reporter assay, and ERK activation results, it was not demonstrated, via in vitro assay, that glucose binds BOSS directly. Because a constant supply of glucose is essential for cell survival, it would be also very difficult to determine in an in vitro system whether glucose is a specific agonist (ligand) for BOSS. In yeasts, the nutrient-sensing activity of Gpr1, the first GPCR glucose sensor to be identified, was demonstrated through phenotypic analysis of Gpr1 mutants. Thus far, the direct interaction between Gpr1 and glucose has yet to be demonstrated. Similar examples are seen with the taste-receptor Tas1r3. Similar problems have also been encountered in identifying the ligands of deorphanizing GPCRs, such as an amino acid-sensing GPCR and proton-sensing GCPRs, even though the agonists of these GPCRs are naturally abundant molecules (Kohyama-Koganeya, 2008).

The mechanisms underlying energy homeostasis in Drosophila and mammals are well conserved: In both, circulating sugars and lipid levels are tightly regulated and depend on insulin signaling and counteracting signals such as glucagon or adrenalin. As for insulin-producing tissues, IPCs and pancreatic β-cells have evolved from a common ancestral insulin-producing neuron. In contrast, adipokinetic hormone (AKH), which corresponds to mammalian glucagon and adrenaline, is produced by the corpora cardiaca cells (CCs) of Drosophila. Both DILPs and AKH signaling regulate hemolymph sugar levels and lipid metabolism. As with the mammalian liver and white adipose tissue, the Drosophila fat body acts as a source of circulating hormones, cytokines, glycogen, and lipids that can influence systemic energy balance and glucose homeostasis. Glucose and lipid metabolism systems are impaired in boss mutants, suggesting that BOSS affects the activity of these pathways. This study demonstrated that insulin signaling is impaired in boss-defective mutants by using tGPH, an insulin signaling activity indicator, and by measuring physiological pAKT levels. Furthermore, the phenotypes of boss-defective mutants are similar to those exhibited by flies in which Inr/PI3K signaling is down-regulated: Both types of flies have elevated glucose levels, and the oenocytes of both boss-defective and FB>dInRDN mutants accumulate lipids under feeding conditions. Because dilps mRNA expression and downstream insulin signaling are not affected by the boss mutation, it is proposed that boss-mediated signaling strongly affects insulin processing, including insulin secretion or stability, but not insulin production in vivo. Recently, two DILP2 binding proteins, imaginal morphogenesis protein-Late 2 (lmp-L2) and acid-labile subunit (ALS), were identified as nutritionally controlled suppressors of growth in Drosophila, demonstrating that the regulatory mechanisms of insulin signaling are very complicated and elaborate. These proteins form a circulating trimeric complex with DILP2 and control animal growth and carbohydrate and fat metabolism. Further analysis is required to understand precisely how and where BOSS signaling regulates insulin-dependent signaling activities (Kohyama-Koganeya, 2008).

Both AKH- and Brummer (BMM) adipose triglyceride lipase-dependent pathways have been reported to be essential for adjusting normal body fat content. AKH and BMM signaling pathways control lipolytic rate, and the lack of dual lipolytic control blocks fat mobilization. Although AKH and BMM signaling activities have not been examined in the boss mutant fat body, it was observed that boss1 mutants have increased hemolymph trehalose levels and sensitivity to starvation stress, indicating that BOSS signaling pathways might also affect AKH and BMM signaling in addition to Inr/PI3K signaling. BOSS-mediated regulation of AKH and BMM may possibly occur through Inr/PI3K signaling, because the cross-talk between insulin signaling and glucagon signaling occurs in mammals and may also occur in Drosophila, as can be deduced from the projection pattern of IPC axons to CCs. Whatever the case, further studies on how BOSS regulates these signaling pathways should provide additional insights into how energy homeostasis is maintained (Kohyama-Koganeya, 2008).

Glucose-sensing pathways trigger metabolic signaling cascades that regulate various aspects of fuel and energy metabolism and influence cell growth, proliferation, and survival. Current knowledge about the upstream sensing mechanisms in glucose regulatory pathways is limited. In mammals, a sugar-sensing GPCR similar to the yeast GPCR that acts through the cAMP pathway has been proposed to exist in intestinal epithelial cells. Furthermore, hypothalamic orexin neurons have been proposed to harbor cell surface receptors that respond to extracellular glucose. However, the precise characteristics of such receptors remain unclear. Because Drosophila shares most of the same basic metabolic functions found in vertebrates and also possesses BOSS orthologous GPCRs (GPRC5Bs), THE identification of a glucose-responding membrane receptor in Drosophila can provide another clue toward deciphering glucose-sensing mechanisms in vertebrates (Kohyama-Koganeya, 2008).


GENE STRUCTURE

Exons - 6

Base pairs in 3' UTR - 213


PROTEIN STRUCTURE

Amino Acids - 896

Structural Domains

The boss gene encodes a protein of 896 amino acids with a putative amino-terminal signal sequence, a large extracellular region of 498 amino acids, and seven potential transmembrane domains followed by a carboxy-terminal cytoplasmic tail of 115 amino acids. The putative membrane localization of the BOSS protein is consistent with a model in which direct interaction between the BOSS and Sevenless proteins specify R7 cell fate. No homology to known G protein-linked receptors was found (Hart, 1990).


bride of sevenless: Regulation | Developmental Biology | Effects of Mutation | References

date revised: 20 June 98

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