title: The carboxy-terminal serine 392 phosphorylation site of human p53 is not required for wild-type activities.
abstract: Wild-type p53 functions in the G1 DNA damage checkpoint pathway by activating gene transcription and preventing cell cycle progression. Others reported that mutation of the serine 386 codon in mouse p53 abolished its ability to suppress growth. Serine 386 of murine p53 and the homologous residue of human p53, serine 392, are phosphorylated in vivo and can be phosphorylated in vitro by casein kinase II (CKII). We constructed mutants that changed serine 392 of human p53 to alanine (p53-S392A) or aspartic acid (p53-S392D); cotransfection of both these mutants with a reporter gene carrying a p53-responsive element into the p53-null Saos-2 cell line activated transcription as well as did wild-type p53. Furthermore, both mutants blocked cell cycle progression after transient transfection in these cells. A stable derivative of the T98G human glioblastoma cell line was established that expressed p53-S392A in response to dexamethasone. Overexpression of this mutant activated transcription of the endogenous waf1 (also called cip1) and mdm2 genes to the same extent as wild-type p53 and also produced growth arrest. Finally, p53-S392A and p53-S392D suppressed foci formation by activated ras and adenovirus E1A oncogenes as efficiently as did wild-type p53. Thus, unlike mutants that altered the serine 15 phosphorylation site, elimination of the serine 392 phosphorylation site had no discernible effect on p53 function. We conclude that neither phosphorylation nor RNA attachment to serine 392 are required for human p53's ability to suppress cell growth or to activate transcription in vivo.
authors.affiliation: Laboratory of Cell Biology, National Institute of Health, Bethesda, Maryland 20892.
authors.forename: M
authors.initials: M
authors.lastname: Fiscella
authors.forename: N
authors.initials: N
authors.lastname: Zambrano
authors.forename: S J
authors.initials: SJ
authors.lastname: Ullrich
authors.forename: T
authors.initials: T
authors.lastname: Unger
authors.forename: D
authors.initials: D
authors.lastname: Lin
authors.forename: B
authors.initials: B
authors.lastname: Cho
authors.forename: W E
authors.initials: WE
authors.lastname: Mercer
authors.forename: C W
authors.initials: CW
authors.lastname: Anderson
authors.forename: E
authors.initials: E
authors.lastname: Appella
mesh_terms: D017500:Adenovirus E1A Proteins / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D047390:Casein Kinase II; D002453:Cell Cycle / Q000235:genetics; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins / Q000235:genetics; D004247:DNA / Q000378:metabolism; D011905:Genes, ras; D006801:Humans; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics; D009687:Nuclear Proteins; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins; D051736:Proto-Oncogene Proteins c-mdm2; D051381:Rats; D011916:Rats, Inbred F344; D012694:Serine / Q000378:metabolism*; D015533:Transcriptional Activation; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*
chemical_list: D017500:Adenovirus E1A Proteins; C493956:CDKN1A protein, human; C493958:Cdkn1a protein, rat; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016213:Cyclins; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D047428:Protein Kinase Inhibitors; D011518:Proto-Oncogene Proteins; D016159:Tumor Suppressor Protein p53; D012694:Serine; D004247:DNA; C497206:MDM2 protein, human; C497208:Mdm2 protein, rat; D051736:Proto-Oncogene Proteins c-mdm2; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases