Accession Number : ADA323678

Title :   General Methods for Identifying G1-Phase Substrates of Cdk Protein Kinases.

Descriptive Note : Annual rept. 1 Jun 95-31 May 96,

Corporate Author : BIOLOGICAL LAB COLD SPRING HARBOR NY

Personal Author(s) : Futcher, A. B. ; Marshak, Daniel R.

PDF Url : ADA323678

Report Date : JUN 1996

Pagination or Media Count : 12

Abstract : We are using two-dimensional gels in combination with genetic and biochemical techniques to identify G1-phase substrates of cyclin-Cdk complexes. We have optimized protocols for labeling proteins with 32P prior to two-dimensional gel electrophoresis. So far, we have successfully identified one important in vivo substrate, Sic1. We have characterized Sic1, and have shown that phosphorylation of Sic1 by cyclin-Cdk complexes promotes DNA synthesis in yeast. We have made peptide antigens for the purpose of making antibodies against phosphorylated Cdk substrates. Mice have been immunized; their sera are positive; and fusions for monoclonal antibodies are about to be done.

Descriptors :   *PHOSPHORUS TRANSFERASES, *BREAST CANCER, PEPTIDES, DEOXYRIBONUCLEIC ACIDS, MICE, MONOCLONAL ANTIBODIES, IN VIVO ANALYSIS, GENETICS, ANTIGENS, ELECTROPHORESIS, BIOSYNTHESIS, SILICA GELS, PHOSPHORYLATION, SERUM GLOBULIN.

Subject Categories : Biochemistry
      Medicine and Medical Research

Distribution Statement : APPROVED FOR PUBLIC RELEASE