Our broad objective is to trace the pathways utilized by receptor tyrosine kinases to regulate cell proliferation and to understand how these pathways are altered in malignancy. Previous studies, from this and other laboratories, have shown that specific cell proteins bind to phosphorylated tyrosine residues in activated receptor tyrosine kinases. The bound proteins regulate different signaling pathways. One pathway leads to the activation of the membrane-associated GTPase, Ras, by promoting the release of GDP from Ras and its replacement by GTP. In its GTP state, Ras binds protein kinases of the Raf family. The membrane-associated Ras-Raf complex is a substrate for one or more other regulatory inputs that together stimulate Raf kinase activity. One known substrate of Raf is responsible for MAP kinase activation, but Raf may have other substrates. We have identified three other proteins capable of specific interactions with RasGTP. These proteins are: a Ral guanine nucleotide dissociation stimulator (GDS), a protein related to the RalGDS, and a protein related to the catalytic (P110) subunits of phosphoinositide 3-kinases. We propose to confirm that these proteins interact with Ras GTP in the cell, determine the biochemical functions of these proteins, and evaluate their roles in oncogenic transformation by activated mutants of Ras. Another goal of this proposal is to identify residues on Ras and Raf involved in binding and generate tools for selective activation of the Raf pathway and other Ras-dependent pathways.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA054786-09
Application #
2894887
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Program Officer
Spalholz, Barbara A
Project Start
1995-09-26
Project End
2001-06-30
Budget Start
1999-07-01
Budget End
2001-06-30
Support Year
9
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Fred Hutchinson Cancer Research Center
Department
Type
DUNS #
075524595
City
Seattle
State
WA
Country
United States
Zip Code
98109
Cary, Leslie A; Klinghoffer, Richard A; Sachsenmaier, Christoph et al. (2002) SRC catalytic but not scaffolding function is needed for integrin-regulated tyrosine phosphorylation, cell migration, and cell spreading. Mol Cell Biol 22:2427-40
Sachsenmaier, C; Sadowski, H B; Cooper, J A (1999) STAT activation by the PDGF receptor requires juxtamembrane phosphorylation sites but not Src tyrosine kinase activation. Oncogene 18:3583-92
Klinghoffer, R A; Sachsenmaier, C; Cooper, J A et al. (1999) Src family kinases are required for integrin but not PDGFR signal transduction. EMBO J 18:2459-71
Kawakami, Y; Hartman, S E; Holland, P M et al. (1998) Multiple signaling pathways for the activation of JNK in mast cells: involvement of Bruton's tyrosine kinase, protein kinase C, and JNK kinases, SEK1 and MKK7. J Immunol 161:1795-802
Winkler, D G; Cutler Jr, R E; Drugan, J K et al. (1998) Identification of residues in the cysteine-rich domain of Raf-1 that control Ras binding and Raf-1 activity. J Biol Chem 273:21578-84
Megidish, T; Cooper, J; Zhang, L et al. (1998) A novel sphingosine-dependent protein kinase (SDK1) specifically phosphorylates certain isoforms of 14-3-3 protein. J Biol Chem 273:21834-45
Gotoh, Y; Cooper, J A (1998) Reactive oxygen species- and dimerization-induced activation of apoptosis signal-regulating kinase 1 in tumor necrosis factor-alpha signal transduction. J Biol Chem 273:17477-82
Winkler, D G; Johnson, J C; Cooper, J A et al. (1997) Identification and characterization of mutations in Ha-Ras that selectively decrease binding to cRaf-1. J Biol Chem 272:24402-9
Finch, A; Holland, P; Cooper, J et al. (1997) Selective activation of JNK/SAPK by interleukin-1 in rabbit liver is mediated by MKK7. FEBS Lett 418:144-8
Chantry, D; Vojtek, A; Kashishian, A et al. (1997) p110delta, a novel phosphatidylinositol 3-kinase catalytic subunit that associates with p85 and is expressed predominantly in leukocytes. J Biol Chem 272:19236-41

Showing the most recent 10 out of 19 publications