A family of related proteins is encoded by the major Drosophila heat shock gene family.

At least four proteins of 70,000 to 75,000 molecular weight (70-75K) were synthesized from mRNA which hybridized with a cloned heat shock gene previously shown to be localized to the 87A and 87C heat shock puff sites. These in vitro-synthesized proteins were indistinguishable from in vivo-synthesized heat shock-induced proteins when analyzed on sodium dodecyl sulfate-polyacrylamide gels. A comparison of the pattern of this group of proteins synthesized in vivo during a 5-min pulse or during continuous labeling indicates that the 72-75K proteins are probably not kinetic precursors to the major 70K heat shock protein. Partial digestion products generated with V8 protease indicated that the 70-75K heat shock proteins are closely related, but that there are clear differences between them. The partial digestion patterns obtained from heat shock proteins from the Kc cell line and from the Oregon R strain of Drosophila melanogaster are very similar. Genetic analysis of the patterns of 70-75K heat shock protein synthesis indicated that the genes encoding at least two of the three 72-75K heat shock proteins are located outside of the major 87A and 87C puff sites.     

Myoblast diversification and ectodermal signaling in Drosophila.

The flight muscles of Drosophila derive from myoblasts found on the third instar disc. We demonstrate that these myoblasts already show distinctive properties and examine how this diversity is generated. In the late larva, Vestigial and low levels of Cut are expressed in myoblasts that will contribute to the indirect flight muscles. Other myoblasts, which express high levels of Cut but no Vestigial, are required for the formation of the direct flight muscles. Vestigial and Cut expression are stabilized by a mutually repressive feedback loop. Vestigial expression begins in the embryo in a subset of adult myoblasts, and Wingless signaling is required later to maintain this expression. Thus, myoblasts are divided into identifiable populations, consistent with their allocation to different muscles, and ectodermal signals act to maintain these differences.     

A large family of divergent Drosophila odorant-binding proteins expressed in gustatory and olfactory sensilla.

We identified a large family of putative odorant-binding protein (OBP) genes in the genome of Drosophila melanogaster. Some of these genes are present in large clusters in the genome. Most members are expressed in various taste organs, including gustatory sensilla in the labellum, the pharyngeal labral sense organ, dorsal and ventral cibarial organs, as well as taste bristles located on the wings and tarsi. Some of the gustatory OBPs are expressed exclusively in taste organs, but most are expressed in both olfactory and gustatory sensilla. Multiple binding proteins can be coexpressed in the same gustatory sensillum. Cells in the tarsi that express OBPs are required for normal chemosensation mediated through the leg, as ablation of these cells dramatically reduces the sensitivity of the proboscis extension reflex to sucrose. Finally, we show that OBP genes expressed in the pharyngeal taste sensilla are still expressed in the poxneuro genetic background while OBPs expressed in the labellum are not. These findings support a broad role for members of the OBP family in gustation and olfaction and suggest that poxneuro is required for cell fate determination of labellar but not pharyngeal taste organs.     

Homology of Drosophila yolk proteins and the triacylglycerol lipase family

Drosophila yolk proteins consist of a set of related proteins of 50,000 Mr. They are derived from slightly larger precursors by cleavage of a signal peptide. In this respect, they differ from the yolk proteins of other insects which are proteolytic fragments of precursors of 200,000 Mr or larger, termed vitellogenins and probably homologous to the vitellogenins of other egg-laying species. We report here a comparative amino acid analysis demonstrating that the Drosophila yolk proteins are non-homologous to the vitellogenin group of yolk proteins, but surprisingly are related to the triacylglycerol lipase family.     

Inhibition of neural crest cell differentiation by embryo ectodermal extract.

The white mutation in Mexican axolotls has long been thought to be a defect associated with the embryonic extracellular environment, but not with embryonic neural crest cells. Thus it was believed that pigment cells in white axolotls disappear from the skin during early development, not because they are intrinsically defective but because they have no choice but to move into an unfavorable environment. We present evidence to suggest that: (1) white neural crest cells are in fact intrinsically different from dark (wild-type) cells, and (2) an inhibitor is produced in white embryonic ectoderm that actively suppresses the migration, differentiation, and survival of pigment cells in this animal. How these observations fit into the existing body of literature on the white mutant and a model for how the white phenotype might develop are discussed.     

The role of Rho family proteins in controlling the migration of crawling cells

Migration of crawling cells (amoebae and some kinds of the tissue cells) is a process related to the dynamic reorganization of actomyosin cytoskeleton. That reorganization engages actin polymerization and de-polymerization, branching of actin network and interaction of myosin II with actin filaments. All those cytoskeleton changes lead to the cell progression, contraction and shifting of the uropod and the cell adhesion. Numerous external stimuli, which activate various surface receptors and signal transduction pathways, can promote migration. Rho family proteins play an important role in the regulation of actin cytoskeleton organization. The most known members of this family are Rho, Rac and Cdc42 proteins, present in all mammalian tissue cells. These proteins control three different stages of cell migration: progression of the frontal edge, adhesion which stabilizes the frontal area, and de-adhesion and shifting of the uropod. Cdc42 and Rac control cell polarization, lamellipodium formation and expansion, organization of focal complexes. Rho protein regulates contractile activity of actomyosin cytoskeleton outside the frontal area, and thus contraction and de-adhesion of the uropod.     

Expression of laminin and of a laminin-related antigen during early development of Drosophila melanogaster

This paper reports the characterization of two immunologically related proteins that may be involved in cell adhesion during Drosophila development. These proteins, laminin chain A and a 240K component, share the epitope recognized by monoclonal antibody RD3 (Mab RD3). The two antigens show different developmental expression profiles. Laminin is detected only from 6 to 8 h of development onwards; its concentration increases during embryogenesis to reach steady-state value in larvae, pupae and adult flies. By contrast, the 240K antigen, not found in oocytes, is present before blastoderm stages; its concentration increases during gastrulation, decreases at the end of organogenesis and the antigen is no longer detected in third instar larvae. Light and electron microscope immunolocalization in imaginal discs indicates that laminin is distributed apically in the lumen and basally in the basal membrane that surrounds the nonevaginated disc. During morphogenesis laminin is detected at the basal side of the evaginating part of the disc epithelium. Immunolocalization on paraffin sections of early embryos suggests that the 240K antigen is related to (1) cell formation and polarization in association with cytoskeleton components, (2) establishment of cell-extracellular substratum interactions during the blastoderm cell sheet organization and (3) basement membrane deposition during embryonic germ cell layer segregation. This 240K protein is poorly or not glycosylated, is resistant to chondroitinase ABC and collagenase and appears therefore as a new extracellular component that might be specifically involved in early processes of morphogenesis.     

A new family of plant antifungal proteins.

Plant seeds contain high concentrations of many antimicrobial proteins. These include chitinases, beta-1,3-glucanases, proteinase inhibitors, and ribosome-inactivating proteins. We recently reported the presence in corn seeds of zeamatin, a protein that has potent activity against a variety of fungi but has none of the above activities. Zeamatin is a 22-kDa protein that acts by causing membrane permeabilization Using a novel bioautography technique, we found similar antifungal proteins in the seeds of 6 of 12 plants examined. A polyclonal antiserum was raised against zeamatin and was used in immunoblots to confirm the presence of zeamatinlike proteins in these seeds. N-terminal amino acid sequencing was carried out on the antifungal proteins from corn, oats, sorghum, and wheat, and these sequences revealed considerable homology with each other. Interestingly, these N-terminal sequences are also similar to those of thaumatin, a pathogenesis-related protein from tobacco, and two salt stress-induced proteins. These results indicate that zeamatin is not unique but is a member of a previously unrecognized family of plant defense proteins that may include some species of pathogenesis-related proteins.