Abstract

The biochemical basis of cellular transformation by the avian sarcoma viruses Rous sarcoma virus, Y73 virus and PRCII virus will be studied. Particular attention will be paid to the role of the phosphorylation of cellular proteins on tyrosine. The role of 1) the phosphorylation of the cytoskeletal protein vinculin and 2) the abundance of fibronectin in the determination of the morphology of transformed cells will be evaluated using viruses which induce atypical morphological transformation. The metabolism and the state of phosphorylation of the two cellular proteins, 80K and 50K, which are found bound to the transforming protein of Rous sarcoma virus will be studied in uninfected cells, in cells transformed by Rous sarcoma virus, and in cells transformed by other agents. Antisera specific to proteins which are substrates of tyrosine protein kinases will be prepared and used to identify cellular proteins which are regulated by the phosphorylation of tyrosine. finally, the state of phosphorylation and the activity of cellular tyrosine protein kinases in uninfected cells and in a variety of transformed cells will be determined. Immediate attention will be paid to the cellular homologue of the transforming protein of Rous sarcoma virus. Antisera useful for the study of the cellular homologues of the transforming proteins of Y73 virus and of PRCII virus will be developed. The structure and the mode of replication of the avian coronavirus infections bronchitis virus will be studied. The polypeptides present in the virion will be enumerated, compared in primary structure, and localized in the virion. The mRNA which is translated to yield each viral structural protein will be determined by in vitro translation of fractionated viral mRNAs. The post-translational processing of each protein will be studied by immunoprecipitation of the viral proteins from infected cells. Finally, the transcriptional mechanism which generates the family of subgenomic mRNAs will be characterized by UV target size analysis.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA017289-10
Application #
3164638
Study Section
Virology Study Section (VR)
Project Start
1979-01-01
Project End
1986-12-31
Budget Start
1985-01-01
Budget End
1985-12-31
Support Year
10
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Salk Institute for Biological Studies
Department
Type
DUNS #
005436803
City
La Jolla
State
CA
Country
United States
Zip Code
92037

  Publications

Schulte, Roberta J; Sefton, Bartholomew M (2003) Inhibition of the activity of SRC and Abl tyrosine protein kinases by the binding of the Wiskott-Aldrich syndrome protein. Biochemistry 42:9424-30
Kjellen, Peter; Amdjadi, Kambiz; Lund, Troy C et al. (2002) The herpesvirus saimiri tip484 and tip488 proteins both stimulate lck tyrosine protein kinase activity in vivo and in vitro. Virology 297:281-8
Amdjadi, K; Sefton, B M (2000) Ultraviolet light-induced stimulation of the JNK mitogen-activated protein kinase in the absence of src family tyrosine kinase activation. J Biol Chem 275:22520-5
Yurchak, L K; Hardwick, J S; Amrein, K et al. (1996) Stimulation of phosphorylation of Tyr394 by hydrogen peroxide reactivates biologically inactive, non-membrane-bound forms of Lck. J Biol Chem 271:12549-54
Schulte, R J; Campbell, M A; Fischer, W H et al. (1994) Tyrosine phosphorylation of VCP, the mammalian homologue of the Saccharomyces cerevisiae CDC48 protein, is unusually sensitive to stimulation by sodium vanadate and hydrogen peroxide. J Immunol 153:5465-72
Taddie, J A; Hurley, T R; Hardwick, B S et al. (1994) Activation of B- and T-cells by the cytoplasmic domains of the B-cell antigen receptor proteins Ig-alpha and Ig-beta. J Biol Chem 269:13529-35
Taddie, J A; Hurley, T R; Sefton, B M (1994) B-cell activation by wild type and mutant Ig-beta cytoplasmic domains. Adv Exp Med Biol 365:23-34
Schulte, R J; Campbell, M A; Fischer, W H et al. (1992) Tyrosine phosphorylation of CD22 during B cell activation. Science 258:1001-4
Campbell, M A; Sefton, B M (1992) Association between B-lymphocyte membrane immunoglobulin and multiple members of the Src family of protein tyrosine kinases. Mol Cell Biol 12:2315-21
Luo, K; Hurley, T R; Sefton, B M (1990) Transfer of proteins to membranes facilitates both cyanogen bromide cleavage and two-dimensional proteolytic mapping. Oncogene 5:921-3

Showing the most recent 10 out of 31 publications