The objectives of the research outlined in this proposal are to define the role of transforming gene products (such as pp60-v-src and p21-v-Ha-ras), serum growth factors or tumor promoting phorbol ester in growth control and cell transformation. The interactions of the transforming proteins with cellular membranes will be characterized and attempts made to determine if any cellular membrane components are required for membrane association by using purified protein components and synthetic lipid vesicles in reconstitution experiments. We shall study cellular enzymes that may be instrumental in causing the phenotypic changes displayed by cells in response to these transforming proteins or tumor promoters. Examples of such cellular enzymes include the serine-specific ribosomal protein S6 kinase, enzymes involved in the regulation of cyclic AMP levels and enzymes involved in the generation of the second messengers diacylglycerol and inositol trisphosphate. When feasible, these cellular enzymes will be used to generate polyclonal or monoclonal antibody as an additional aid in functional characterization and for use in obtaining cDNA or genomic molecular clones for these proteins. We shall compare quiescent cells to cells stimulated to proliferate with regard to synthesis of specific proteins, protein phosphorylation patterns and expression of cellular genes. These studies should provide new insights concerning the biochemical pathways that lead to cellular proliferation and malignant transformation. We shall utilize 1) multiple techniques for protein purification; 2) molecular cloning of cDNA libraries; 3) polyclonal and monoclonal antibodies; 4) viral transformation mutants.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Unknown (R35)
Project #
5R35CA042580-02
Application #
3479462
Study Section
(SRC)
Project Start
1986-07-01
Project End
1993-06-30
Budget Start
1987-07-01
Budget End
1988-06-30
Support Year
2
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Harvard University
Department
Type
Schools of Arts and Sciences
DUNS #
071723621
City
Cambridge
State
MA
Country
United States
Zip Code
02138
Ma, Sheng; Liu, Mei-Ann; Yuan, Yi-Lu O et al. (2003) The serum-inducible protein kinase Snk is a G1 phase polo-like kinase that is inhibited by the calcium- and integrin-binding protein CIB. Mol Cancer Res 1:376-84
Lin, C Y; Madsen, M L; Yarm, F R et al. (2000) Peripheral Golgi protein GRASP65 is a target of mitotic polo-like kinase (Plk) and Cdc2. Proc Natl Acad Sci U S A 97:12589-94
Song, S; Grenfell, T Z; Garfield, S et al. (2000) Essential function of the polo box of Cdc5 in subcellular localization and induction of cytokinetic structures. Mol Cell Biol 20:286-98
Gallina, A; Simoncini, L; Garbelli, S et al. (1999) Polo-like kinase 1 as a target for human cytomegalovirus pp65 lower matrix protein. J Virol 73:1468-78
Lee, K S; Song, S; Erikson, R L (1999) The polo-box-dependent induction of ectopic septal structures by a mammalian polo kinase, plk, in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A 96:14360-5
Greulich, H; Erikson, R L (1998) An analysis of Mek1 signaling in cell proliferation and transformation. J Biol Chem 273:13280-8
Brott, B K; Pinsky, B A; Erikson, R L (1998) Nlk is a murine protein kinase related to Erk/MAP kinases and localized in the nucleus. Proc Natl Acad Sci U S A 95:963-8
Lee, K S; Grenfell, T Z; Yarm, F R et al. (1998) Mutation of the polo-box disrupts localization and mitotic functions of the mammalian polo kinase Plk. Proc Natl Acad Sci U S A 95:9301-6
Alessandrini, A; Chiaur, D S; Pagano, M (1997) Regulation of the cyclin-dependent kinase inhibitor p27 by degradation and phosphorylation. Leukemia 11:342-5
Alessandrini, A; Brott, B K; Erikson, R L (1997) Differential expression of MEK1 and MEK2 during mouse development. Cell Growth Differ 8:505-11

Showing the most recent 10 out of 50 publications