This project is designed to understand the role of tyrosine protein phosphorylation in the activation of B lymphocytes by antigen. The binding of antigen or anti-immunoglobulin antibody to immunoglobulin M or D on the surface of mature B cells induces the rapid phosphorylation of approximately 10 proteins on tyrosine. Surface immunoglobulin therefore must be coupled in some manner to one or more tyrosine protein kinases or phosphatases. Because an inhibitor of tyrosine phosphorylation renders B cells unresponsive to stimulation through the antigen receptor, tyrosine phosphorylation is apparently important in B cell activation. B cells that do not express the tyrosine protein phosphatase CD45 are unresponsive to the cross-linking of surface immunoglobulin. This insensitivity could be due to increased inhibitory phosphorylation of a tyrosine protein kinase. We will therefore study the phosphorylation of the several src-family tyrosine protein kinases found associated with the antigen receptor complex. Additionally, we will ask whether the insensitivity of CD45-negative B cells is reversed by introduction into them of a constitutively-activated version of a src-family kinase. We will also study the interaction of the src-family kinases with the antigen receptor complex. We will identify the domain of the src kinases responsible for their interaction by introduction into B cells of mutated src kinases. Additionally, we will ask whether the cytoplasmic domains of either or both of the immunoglobulin-associated proteins, Ig-alpha and Ig-beta, can interact with tyrosine kinases directly. If so, we will ask whether these domains can participate in signal transduction. Finally, we will exploit newly-isolated monoclonal antibodies to two novel tyrosine phosphorylated proteins in activated B cells. The 95K and 140K proteins that these antibodies recognize will be characterized and their genes cloned. These studies have the potential to reveal the identities of tyrosine protein kinases and substrates that are important in the regulation of the proliferation of normal B cells and may play a role in the unregulated growth of malignant B cells such as those transformed by Abelson murine leukemia virus.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA017289-26
Application #
6350011
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1979-01-01
Project End
2003-01-31
Budget Start
2001-02-01
Budget End
2003-01-31
Support Year
26
Fiscal Year
2001
Total Cost
$395,207
Indirect Cost
Name
Salk Institute for Biological Studies
Department
Type
DUNS #
005436803
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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