title: Excess HDM2 impacts cell cycle and apoptosis and has a selective effect on p53-dependent transcription.
abstract: Mutational inactivation of p53 is only one of the ways that tumors lose p53 function. An alternate route is through overexpression of HDM2, the negative regulator of p53. To further understand how excess HDM2 regulates p53-mediated functions, we generated H1299 cell clones that constitutively express both ectopic HDM2 and tetracycline-regulated inducible p53. We found that over a range of p53 concentrations constitutively expressed HDM2 did not affect the levels of p53 protein. Nevertheless, cells with excess HDM2 displayed numerous changes in their response to p53. After DNA damage, such cells had both increased p53-mediated G2 arrest and reduced cell death. They also showed selective impairment of p53 target gene induction in that some p53 targets were unaffected whereas others were markedly less well induced in the presence of extra HDM2 protein. We also found that excess HDM2 was correlated with reduced p53 acetylation but did not affect p53 association with target promoters in vivo. Indeed, there was no significant difference in the amount of HDM2 associated with p53 at target promoters that differed in their expression depending on the presence of extra HDM2. Thus, HDM2 can selectively down-regulate the transcription function of p53 without either degrading p53 or affecting the interaction of p53 with target promoters.
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York 10027; Cancer Research Laboratory, Taiho Pharmaceutical Co., Ltd., 1-27 Misugidai, Hanno, Saitama 357-8527, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Ohkubo
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York 10027.
authors.forename: Tomoaki
authors.initials: T
authors.lastname: Tanaka
authors.affiliation: National Cancer Research Institute, 5-1-1 Tsukiji, Chuo-ku, Tokyo 104-0045, Japan.
authors.forename: Yoichi
authors.initials: Y
authors.lastname: Taya
authors.affiliation: Cancer Research Laboratory, Taiho Pharmaceutical Co., Ltd., 1-27 Misugidai, Hanno, Saitama 357-8527, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Kitazato
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York 10027. Electronic address: clp3@columbia.edu.
authors.forename: Carol
authors.initials: C
authors.lastname: Prives
mesh_terms: D017209:Apoptosis; D002453:Cell Cycle; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis; D015536:Down-Regulation; D015972:Gene Expression Regulation, Neoplastic; D016158:Genes, p53; D056284:Histone Deacetylase 1; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D011401:Promoter Regions, Genetic; D051736:Proto-Oncogene Proteins c-mdm2 / Q000737:chemistry / Q000502:physiology*; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*
chemical_list: D016159:Tumor Suppressor Protein p53; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; C110550:HDAC1 protein, human; D056284:Histone Deacetylase 1; D006655:Histone Deacetylases