Inactivation of both alleles of the fruit¯y D. melanogaster brain tumor (brat) gene results in the production of a tumor-like neoplasm in the larval brain, and lethality in the larval third instar and pupal stages. We cloned the brat gene from a transposon-tagged allele and identi®ed its gene product. brat encodes for an 1037 amino acid protein with an N-terminal B-box1 zinc ®nger followed by a B-box2 zinc ®nger, a coiled-coil domain, and a C-terminal b-propeller domain with six blades. All these motifs are known to mediate protein ± protein interactions. Sequence analysis of four brat alleles revealed that all of them are mutated at the bpropeller domain. The clustering of mutations in this domain strongly suggests that it has a crucial role in the normal function of Brat, and de®nes a novel protein motif involved in tumor suppression activity. The brat gene is expressed in the embryonic central and peripheral nervous systems including the embryonic brain. In third instar larva brat expression was detected in the larval central nervous system including the brain and the ventral ganglion, in two glands ± the ring gland and the salivary gland, and in parts of the foregut ± the gastric caecae and the proventriculus. A second brat-like gene was found in D. melanogaster, and homologs were identi®ed in the nematode, mouse, rat, and human. Accumulated data suggests that Brat may regulate proliferation and di erentiation by secretion/transportmediated processes.
The transcription patterns of three v-Ha-ras-related cellular oncogenes in Drosophila melanogaster were studied. Each gene coded for at least two distinct transcripts. The larger transcript of each gene was expressed at a similar abundance during the entire life cycle of fruit flies, whereas the shorter transcripts were much more abundant in embryonic stages than at later stages.
The promoter of the Drosophila melanogaster Ras2 gene is bidirectional, regulating an additional gene oriented in the opposite polarity. The two divergently transcribed genes are only 93 bases apart and deletion analysis proved that common cis-acting elements within this promoter region are required for the transcriptional activity of both genes. We cloned the gene paired with Ras2 in the bidirectional promoter and isolated cDNAs corresponding to its mRNA. The Ras opposite (Rop) gene encodes for a 68 × 10(3) M(r) protein which shares sequence homology with the members of a novel Saccharomyces cerevisiae gene family, including the SLY1, SEC1 and VPS33 (SLP1) genes, all of which are involved in vesicle trafficking among yeast cellular compartments. A highly conserved motif in this family is also found in beta-COP, a coat protein isolated from rat Golgi-bound nonclathrin vesicles. Thus, the Rop protein may be a component of one of the vesicle trafficking pathways in Drosophila cells. The Rop gene expression during embryogenesis is restricted to the central nervous system (CNS) and the garland cells, a small group of nephrocytes that takes up waste materials from the haemolymph by endocytosis. Ras2 is also expressed in the embryonic garland cells. In postembryonic stages, the two genes are co-expressed in the larval salivary glands and the central nervous system, and in the adult CNS and reproductive systems. Interestingly, the S. cerevisiae SLY1-20 allele is a suppressor of the loss of the YPT1 gene, a ras-like gene implicated in vesicle translocation, suggesting that the two genes may interact with one another. Since Sec1p and beta-COP may also interact with small GTP-binding proteins of the ras superfamily, it is conceivable that the Rop and Ras2 gene products are not just co-expressed in common tissues, but may also functionally interact with one another in these tissues.
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