Evidence for interaction between v-Mos and a p34cdc2 isoform, p35cdk.
The c-mos gene product (c-Mos) encodes a serine/threonine kinase required for activation of pre-MPF (maturation-promoting factor) to MPF in oocytes undergoing meiosis and also for stabilization of MPF leading to metaphase arrest in unfertilized eggs. In order to determine whether the v-mos gene product (v-Mos) causes neoplastic transformation via interaction with cell cycle control elements, we have searched for proteins that interact with v-Mos. Extracts of NIH3T3 cells transformed by v-Mos encoded by Moloney murine sarcoma virus (Mo-MuSV) were examined by gel filtration, by immunoprecipitation with antibodies to a conserved region of p34cdc2, and by binding to beads that contain cross-linked p13suc1, a protein known to bind p34cdc2. Gel filtration detected a 500-kDa complex that contained v-Mos and a p34cdc2 isoform, termed p35cdk. The 500-kDa macromolecular complex also exhibited histone H1 phosphorylation activity, consistent with the presence of a cdc2 isoform. The identity of p35cdk is based on its recognition by anti-cdc2 PSTAIR but not by anti-cdc2 C-terminal antibodies, which detect authentic p34cdc2. Structures containing v-Mos and p35cdk were also detected by experiments involving co-immunoprecipitation of v-Mos with anti-cdc2 PSTAIR antibodies. Furthermore, both v-Mos and the p35cdk co-precipitated with p13suc1-Sepharose beads. Our findings raise the possibility of a v-Mos-p35cdk regulatory interaction in cells transformed by Mo-MuSV.
Bai
W
W
Department of Molecular Pathology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Singh
B
B
Yang
Y
Y
Arlinghaus
R B
RB
D016475:3T3 Cells; D000595:Amino Acid Sequence; D000818:Animals; D002453:Cell Cycle; D002471:Cell Transformation, Neoplastic; D016200:Maturation-Promoting Factor / Q000378:metabolism; D000090204:Mesothelin; D051379:Mice; D008969:Molecular Sequence Data; D017232:Oncogene Proteins v-mos; D011233:Precipitin Tests; D006656:Protamine Kinase / Q000032:analysis; D011494:Protein Kinases / Q000032:analysis*; D017346:Protein Serine-Threonine Kinases; D015682:Retroviridae Proteins, Oncogenic / Q000032:analysis* / Q000633:toxicity
C000715528:Msln protein, mouse; D017232:Oncogene Proteins v-mos; D015682:Retroviridae Proteins, Oncogenic; D011494:Protein Kinases; D006656:Protamine Kinase; D017346:Protein Serine-Threonine Kinases; D016200:Maturation-Promoting Factor; D000090204:Mesothelin