The function of the ras family of oncogene proteins will be investigated utilizing both living cell systems and in vitro biochemical approaches. The human H-ras proto- and oncogene proteins have been produced in an E. coli expression system, and the proteins have been purified to homogeneity and characterized as """"""""native"""""""" entities. We propose to use these intact proteins as antigens for the production of a bank of mouse-derived monoclonal antibodies, which will be subsequently employed in both biochemical experiments and microinjection experiments with normal and transformed cells. As we have recently shown, microinjection of the human H-ras oncogene protein, but not the proto-oncogene protein, into quiescent normal cells results in rapid morphological changes followed by DNA synthesis and proliferation. We wish to follow up these initial experiments by examining this unique assay of the transformation process induced by human oncogene proteins in great detail. And, through the use of the purified human H-ras proteins, we intend to look at the interactions of other cellular proteins with the ras proteins in vitro. (X)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA039811-01
Application #
3179202
Study Section
Molecular Biology Study Section (MBY)
Project Start
1985-04-01
Project End
1988-03-31
Budget Start
1985-04-01
Budget End
1986-03-31
Support Year
1
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Cold Spring Harbor Laboratory
Department
Type
DUNS #
065968786
City
Cold Spring Harbor
State
NY
Country
United States
Zip Code
11724
LaMorte, V J; Kennedy, E D; Collins, L R et al. (1993) A requirement for Ras protein function in thrombin-stimulated mitogenesis in astrocytoma cells. J Biol Chem 268:19411-5
Sosnowski, R G; Feldman, S; Feramisco, J R (1993) Interference with endogenous ras function inhibits cellular responses to wounding. J Cell Biol 121:113-9
Walton, P A; Gould, S J; Rachubinski, R A et al. (1992) Transport of microinjected alcohol oxidase from Pichia pastoris into vesicles in mammalian cells: involvement of the peroxisomal targeting signal. J Cell Biol 118:499-508
Lin, A; Frost, J; Deng, T et al. (1992) Casein kinase II is a negative regulator of c-Jun DNA binding and AP-1 activity. Cell 70:777-89
Walton, P A; Gould, S J; Feramisco, J R et al. (1992) Transport of microinjected proteins into peroxisomes of mammalian cells: inability of Zellweger cell lines to import proteins with the SKL tripeptide peroxisomal targeting signal. Mol Cell Biol 12:531-41
Fantozzi, D A; Taylor, S S; Howard, P W et al. (1992) Effect of the thermostable protein kinase inhibitor on intracellular localization of the catalytic subunit of cAMP-dependent protein kinase. J Biol Chem 267:16824-8
Lamb, N J; Fernandez, A; Feramisco, J R et al. (1989) Modulation of vimentin containing intermediate filament distribution and phosphorylation in living fibroblasts by the cAMP-dependent protein kinase. J Cell Biol 108:2409-22
Lamb, N J; Fernandez, A; Conti, M A et al. (1988) Regulation of actin microfilament integrity in living nonmuscle cells by the cAMP-dependent protein kinase and the myosin light chain kinase. J Cell Biol 106:1955-71
Bar-Sagi, D; Gomperts, B D (1988) Stimulation of exocytotic degranulation by microinjection of the ras oncogene protein into rat mast cells. Oncogene 3:463-9
Bar-Sagi, D; Suhan, J P; McCormick, F et al. (1988) Localization of phospholipase A2 in normal and ras-transformed cells. J Cell Biol 106:1649-58

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