Changes in phosphorylation of myc oncogene and RB antioncogene protein products during growth arrest of the murine lymphoma WEHI 231 cell line.
The expression of the protein products of the c-myc oncogene and retinoblastoma susceptibility gene (RB) was investigated during either goat anti-mouse immunoglobulin (GaMIg)- or phorbol ester (TPA)-induced growth arrest of the murine B-lymphoma cell line WEHI 231. Previously we have demonstrated that c-myc mRNA levels increase within 1-2 h of treatment, return to control levels by 4 h, and decline below these values by 24 h of treatment. Here we demonstrate that the level of c-myc protein synthesis and mRNA change in parallel. The predominant c-myc protein expressed during the time course is the one initiated at the AUG codon (P2). The myc protein synthesized following 1-2 h of anti-immunoglobulin or TPA treatment migrates more slowly in a polyacrylamide gel as a result of increased phosphorylation. This hyperphosphorylation was no longer detectable by 4-6 h of treatment. Furthermore, the hyperphosphorylated myc protein appears to be more readily extractable with salt than the hypophosphorylated form. The product of the RB gene is present in multiple phosphorylation states in exponentially growing WEHI 231 cells. By 8 h of GaMIg or TPA treatment, a hypophosphorylated form begins to be detectable and significant levels were seen by 15 h. Thus post-translational control of both c-myc and RB expression occurs during the growth arrest of WEHI 231 cells. These changes in phosphorylation may play a role in mediating the cessation of proliferation of these cells.
Maheswaran
S
S
Department of Biochemistry, Boston University School of Medicine, Massachusetts 02118.
McCormack
J E
JE
Sonenshein
G E
GE
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002455:Cell Division; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D016259:Genes, myc; D008223:Lymphoma / Q000235:genetics*; D051379:Mice; D010766:Phosphorylation; D014176:Protein Biosynthesis; D016271:Proto-Oncogene Proteins c-myc / Q000187:drug effects / Q000502:physiology*; D012313:RNA / Q000032:analysis; D016160:Retinoblastoma Protein / Q000187:drug effects / Q000502:physiology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D016271:Proto-Oncogene Proteins c-myc; D016160:Retinoblastoma Protein; D012313:RNA; D013755:Tetradecanoylphorbol Acetate