Abstract

The experiments outlined in this proposal are aimed at identifying the function of a proto-oncogene called yes. This cellular gene is highly expressed in cerebellum, retina, kidney and liver. Molecular, biochemical and biological approaches are proposed to determine which cells in these organs and tissues express the yes gene, and to identify cellular functions governed by this gene and its products. Polyclonal antibodies against the viral-yes protein, which recognize cellular-yes (c-yes) proteins, and monoclonal antibodies that recognize avian and rodent yes proteins will be used as probes. Attempts will also be made to map neuronal cells that express c-yes proteins by analyzing mutant in mice in which mutations cause death of one or more classes of cerebellar cells. Efforts will be made to identify and characterize proteins that are phosphorylated on tyrosine specifically in the cells expressing the yes proteins, which are tyrosine protein kinases. As a part of this project we will also analyze the mechanism of activation of the c-yes protein by polyoma middle T antigen and we will study the influence of post-translational modifications of the c-yes protein on its kinase activity. These studies may elucidate the mechanisms involved in regulating the c-yes protein kinase activity in normal cells. Structure of the yes proto-oncogene and the mechanism of c-yes transduction into the Yamaguchi 73 viral genome will be studied by sequence analysis of molecular clones. In addition, characterization of cDNA clones will be carried out to investigate differential splicing of c-yes transcripts. Analysis of the c-yes cDNA expression in a cloned retrovirus vector should provide information on the transforming potential of this proto-oncogene. If the c-yes cDNA expressed in a retroviral context is not transforming, we will map spontaneous mutations that convert the c-yes gene into a transforming gene. Functional and structural analysis of cellular proto-oncogenes may have broad implications for our understanding of the molecular mechanisms of neoplastic transformation and could provide some clues of strategies for controlling them.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29CA045757-04
Application #
3458525
Study Section
Molecular Cytology Study Section (CTY)
Project Start
1987-08-01
Project End
1992-07-31
Budget Start
1990-08-01
Budget End
1991-07-31
Support Year
4
Fiscal Year
1990
Total Cost
Indirect Cost

  Institution

Name
Rockefeller University
Department
Type
Other Domestic Higher Education
DUNS #
071037113
City
New York
State
NY
Country
United States
Zip Code
10065

  Publications

Espanel, X; Sudol, M (2001) Yes-associated protein and p53-binding protein-2 interact through their WW and SH3 domains. J Biol Chem 276:14514-23
Strano, S; Munarriz, E; Rossi, M et al. (2001) Physical interaction with Yes-associated protein enhances p73 transcriptional activity. J Biol Chem 276:15164-73
Martins-Green, M; Bixby, J L; Yamamoto, T et al. (2000) Tissue specific expression of Yrk kinase: implications for differentiation and inflammation. Int J Biochem Cell Biol 32:351-64
Summy, J M; Guappone, A C; Sudol, M et al. (2000) The SH3 and SH2 domains are capable of directing specificity in protein interactions between the non-receptor tyrosine kinases cSrc and cYes. Oncogene 19:155-60
Chang, A; Cheang, S; Espanel, X et al. (2000) Rsp5 WW domains interact directly with the carboxyl-terminal domain of RNA polymerase II. J Biol Chem 275:20562-71
Luton, F; Verges, M; Vaerman, J P et al. (1999) The SRC family protein tyrosine kinase p62yes controls polymeric IgA transcytosis in vivo. Mol Cell 4:627-32
Espanel, X; Sudol, M (1999) A single point mutation in a group I WW domain shifts its specificity to that of group II WW domains. J Biol Chem 274:17284-9
Mohler, P J; Kreda, S M; Boucher, R C et al. (1999) Yes-associated protein 65 localizes p62(c-Yes) to the apical compartment of airway epithelia by association with EBP50. J Cell Biol 147:879-90
Ermekova, K S; Chang, A; Zambrano, N et al. (1998) Proteins implicated in Alzheimer disease. The role of FE65, a new adapter which binds to beta-amyloid precursor protein. Adv Exp Med Biol 446:161-80
Ermekova, K S; Zambrano, N; Linn, H et al. (1997) The WW domain of neural protein FE65 interacts with proline-rich motifs in Mena, the mammalian homolog of Drosophila enabled. J Biol Chem 272:32869-77

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