Basement membranes are thin condensations of specialized extracellular matrix that underlie epithelia, surround muscle cells, and serve various physiological functions. Their pathobiological chemistry is important in several diseases, as well as in both aging and development. Their durability and replacement are not understood. We have shown that several major constituents of basement membranes: procollagen IV, laminin and entactin, are closely similar in man and Drosophila melanogaster, and have established methodology for studying basement membrane formation in Drosophila. We have cDNA clones for these materials and for some others and have obtained the complete nucleic acid sequence of Drosophila procollagen IV. In the lethal mutant, myospheroid, in which muscle basement membranes detach, we found that this gene codes for part of an extracellular cell surface receptor. We shall investigate the molecular functions of those parts of basement membrane components and their receptors which are highly similar in man and Drosophila, for example, the carboxyl (NC1) junctional domain of procollagen IV. We shall use the special advantages of the Drosophila system to reintroduce into the gene line genes that have been changed by specific, site-directed mutagenesis at evolutionary conserved 1 parts of basement membrane proteins. We shall also introduce these modified genes into Drosophila mass cell cultures that secrete basement membrane components, isolate the modified proteins, and study their interactions in vitro. We will complete our ongoing characterization of several high molecular weight glycoproteins that are secreted by Drosophila cells and are putative additional components of basement membranes. The Drosophila receptor chain that we found has a very similar vertebrate homolog, the integrin- beta chain, which is the common component of several important receptors for extracellular matrix molecules. We plan to isolate corresponding Drosophila receptors and characterize them. We have demonstrated the utility of specific antibodies and nucleic acid probes for defining the time and site of appearance of basement membrane collagen IV, and its mRNA, during Drosophila development. We will use similar probes for Drosophila laminin and entactin. We shall fuse the promoter regions of basement membrane genes to the marker beta-galactosidase, to study the action of the promoters, and will search for common controlling elements of basement membrane proteins and their receptors.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
5R01AG002128-09
Application #
3114332
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1979-09-29
Project End
1993-04-30
Budget Start
1989-05-18
Budget End
1990-04-30
Support Year
9
Fiscal Year
1989
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Type
Schools of Arts and Sciences
DUNS #
119132785
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
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Kramerova, I A; Kramerov, A A (1999) Mucinoprotein is a universal constituent of stable intercellular bridges in Drosophila melanogaster germ line and somatic cells. Dev Dyn 216:349-60
Bunch, T A; Graner, M W; Fessler, L I et al. (1998) The PS2 integrin ligand tiggrin is required for proper muscle function in Drosophila. Development 125:1679-89
Mayer, U; Mann, K; Fessler, L I et al. (1997) Drosophila laminin binds to mammalian nidogen and to heparan sulfate proteoglycan. Eur J Biochem 245:745-50
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Parker, C G; Fessler, L I; Nelson, R E et al. (1995) Drosophila UDP-glucose:glycoprotein glucosyltransferase: sequence and characterization of an enzyme that distinguishes between denatured and native proteins. EMBO J 14:1294-303
Murray, M A; Fessler, L I; Palka, J (1995) Changing distributions of extracellular matrix components during early wing morphogenesis in Drosophila. Dev Biol 168:150-65
Gullberg, D; Fessler, L I; Fessler, J H (1994) Differentiation, extracellular matrix synthesis, and integrin assembly by Drosophila embryo cells cultured on vitronectin and laminin substrates. Dev Dyn 199:116-28
Fogerty, F J; Fessler, L I; Bunch, T A et al. (1994) Tiggrin, a novel Drosophila extracellular matrix protein that functions as a ligand for Drosophila alpha PS2 beta PS integrins. Development 120:1747-58

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