The objective of this proposal is to examine the mechanism by which the transforming gene, src, encoded by Rous sarcoma virus alters the expression of normal cellular genes. The model system utilizes chicken embryo chondrogenic cells infected with ts-src mutants of Rous sarcoma virus. We have demonstrated that transformation induces changes in the synthesis of proteoglycans, collagens, fibrorectin, and hyaluronic acid. Here we will focus on the synthesis of the chondroblast specific type II collagen propeptide Alpha I(II) which is switched off by transformation and fibrorectin and the type I collagen propeptides Alpha 1(I) and Alpha 2(I) which are switched on. The levels of analysis include analysis of synthesis of the gene products, synthesis and processing of the specific m-RNA's, and analysis of DNase I sensitivity and methylation of the specific DNA coding regions. The mechanism by which the altered expression is achieved by RSV-induced cell transformation will be compared to (a) changes in gene expression during normal chondrogenic differentiation, (b) other physical and chemical treatments which induce analogous alterations in chondroblasts, and (c) changes induced by src variants, other sarcoma virus strains, and leukemogenic viruses. This comparative approach combined with drug inhibition studies and kinetics of the various transitions will be used to refine and examine the hypotheses on the mechanism of src action.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA016502-11
Application #
3164422
Study Section
Virology Study Section (VR)
Project Start
1983-02-01
Project End
1987-11-30
Budget Start
1984-12-01
Budget End
1985-11-30
Support Year
11
Fiscal Year
1985
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Type
Schools of Medicine
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Boettiger, David (2007) Quantitative measurements of integrin-mediated adhesion to extracellular matrix. Methods Enzymol 426:1-25
Lee, Mark H; Ducheyne, Paul; Lynch, Laura et al. (2006) Effect of biomaterial surface properties on fibronectin-alpha5beta1 integrin interaction and cellular attachment. Biomaterials 27:1907-16
Lynch, Laura; Vodyanik, Pavel I; Boettiger, David et al. (2005) Insulin-like growth factor I controls adhesion strength mediated by alpha5beta1 integrins in motile carcinoma cells. Mol Biol Cell 16:51-63
Lee, Fang-Hua; Haskell, Christopher; Charo, Isreal F et al. (2004) Receptor-ligand binding in the cell-substrate contact zone: a quantitative analysis using CX3CR1 and CXCR1 chemokine receptors. Biochemistry 43:7179-86
Guvakova, Marina A; Boettiger, David; Adams, Josephine C (2002) Induction of fascin spikes in breast cancer cells by activation of the insulin-like growth factor-I receptor. Int J Biochem Cell Biol 34:685-98
Datta, Anirban; Huber, Francois; Boettiger, David (2002) Phosphorylation of beta3 integrin controls ligand binding strength. J Biol Chem 277:3943-9
Garcia, Andres J; Schwarzbauer, Jean E; Boettiger, David (2002) Distinct activation states of alpha5beta1 integrin show differential binding to RGD and synergy domains of fibronectin. Biochemistry 41:9063-9
Boettiger, D; Lynch, L; Blystone, S et al. (2001) Distinct ligand-binding modes for integrin alpha(v)beta(3)-mediated adhesion to fibronectin versus vitronectin. J Biol Chem 276:31684-90
Boettiger, D; Huber, F; Lynch, L et al. (2001) Activation of alpha(v)beta3-vitronectin binding is a multistage process in which increases in bond strength are dependent on Y747 and Y759 in the cytoplasmic domain of beta3. Mol Biol Cell 12:1227-37
Garcia, A J; Boettiger, D (1999) Integrin-fibronectin interactions at the cell-material interface: initial integrin binding and signaling. Biomaterials 20:2427-33

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