A DNA damage-induced p53 serine 392 kinase complex contains CK2, hSpt16, and SSRP1.

Phosphorylation of the human p53 protein at Ser-392 has been shown to be responsive to UV but not gamma irradiation. Here we describe identification and purification of a mammalian UV-activated protein kinase complex that phosphorylates Ser-392 of p53 in vitro. This kinase complex contains casein kinase 2 (CK2) and the chromatin transcriptional elongation factor FACT (a heterodimer of hSpt16 and SSRP1). In vitro studies show that FACT alters the specificity of CK2 in the complex such that it selectively phosphorylates p53 over other substrates including casein. In addition, phosphorylation by the kinase complex enhances p53 activity. These results thus provide a potential mechanism for p53 activation by UV irradiation.

[1]  E. Stavridi,et al.  Phosphorylation of Ser-20 mediates stabilization of human p53 in response to DNA damage. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[2]  M. Scheffner,et al.  Functional domains of wild-type and mutant p53 proteins involved in transcriptional regulation, transdominant inhibition, and transformation suppression , 1993, Molecular and cellular biology.

[3]  O. Issinger,et al.  Stress-induced Activation of Protein Kinase CK2 by Direct Interaction with p38 Mitogen-activated Protein Kinase* , 2000, The Journal of Biological Chemistry.

[4]  M. Ljungman,et al.  Blockage of RNA polymerase as a possible trigger for u.v. light-induced apoptosis. , 1996, Oncogene.

[5]  H. Sakamoto,et al.  Phosphorylation of serine 392 stabilizes the tetramer formation of tumor suppressor protein p53. , 1997, Biochemistry.

[6]  S. Jackson,et al.  Regulation of p53 in response to DNA damage , 1999, Oncogene.

[7]  E. Harlow,et al.  Antibodies: A Laboratory Manual , 1988 .

[8]  S. Lowe,et al.  Stabilization of the p53 tumor suppressor is induced by adenovirus 5 E1A and accompanies apoptosis. , 1993, Genes & development.

[9]  K. Kinzler,et al.  Identification of p53 as a sequence-specific DNA-binding protein , 1991, Science.

[10]  G. Orphanides,et al.  FACT, a Factor that Facilitates Transcript Elongation through Nucleosomes , 1998, Cell.

[11]  E. Wagner,et al.  Down‐regulation of wild‐type p53 activity interferes with apoptosis of IL‐3‐dependent hematopoietic cells following IL‐3 withdrawal. , 1994, The EMBO journal.

[12]  T. Lindahl,et al.  Quality control by DNA repair. , 1999, Science.

[13]  A. Levine,et al.  Ultraviolet radiation, but not gamma radiation or etoposide-induced DNA damage, results in the phosphorylation of the murine p53 protein at serine-389. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  D. Meek,et al.  The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II. , 1990, The EMBO journal.

[15]  A. Levine.  p53, the Cellular Gatekeeper for Growth and Division , 1997, Cell.

[16]  E. Stavridi,et al.  ATM-dependent activation of p53 involves dephosphorylation and association with 14-3-3 proteins , 1998, Nature Genetics.

[17]  J. Darnell,et al.  The inhibition of DRB (5,6-dichloro-1-β-d-ribofuranosylbenzimidazole) of hnRNA and mRNA production in HeLa cells , 1976, Cell.

[18]  M. Kapoor,et al.  The p38MAPK inhibitor SB203580 alleviates ultraviolet-induced phosphorylation at serine 389 but not serine 15 and activation of p53. , 1999, Biochemical and biophysical research communications.

[19]  V. Adler,et al.  p53 phosphorylation and association with murine double minute 2, c-Jun NH2-terminal kinase, p14ARF, and p300/CBP during the cell cycle and after exposure to ultraviolet irradiation. , 2000, Cancer research.

[20]  D. Pim,et al.  Monoclonal antibodies specific for simian virus 40 tumor antigens , 1981, Journal of virology.

[21]  C. N. Coleman,et al.  Increases in sequence specific DNA binding by p53 following treatment with chemotherapeutic and DNA damaging agents. , 1993, Cancer research.

[22]  P. Sharp,et al.  5,6-Dichloro-1-beta-D-ribofuranosylbenzimidazole inhibits transcription elongation by RNA polymerase II in vitro. , 1989, The Journal of biological chemistry.

[23]  S. Elledge,et al.  DNA damage-induced activation of p53 by the checkpoint kinase Chk2. , 2000, Science.

[24]  T. Hupp,et al.  DNA damage triggers DRB-resistant phosphorylation of human p53 at the CK2 site , 1998, Oncogene.

[25]  K. Khanna,et al.  Ionizing radiation and UV induction of p53 protein by different pathways in ataxia-telangiectasia cells. , 1993, Oncogene.

[26]  E. Appella,et al.  Prolonged p53 protein accumulation in trichothiodystrophy fibroblasts dependent on unrepaired pyrimidine dimers on the transcribed strands of cellular genes , 1997, Molecular carcinogenesis.

[27]  A. Levine,et al.  The CDK7-cycH-p36 complex of transcription factor IIH phosphorylates p53, enhancing its sequence-specific DNA binding activity in vitro , 1997, Molecular and cellular biology.

[28]  R. Weinmann,et al.  Casein kinase type II is involved in the inhibition by 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole of specific RNA polymerase II transcription. , 1986, The Journal of biological chemistry.

[29]  M. Kapoor,et al.  Mutation of Phosphoserine 389 Affects p53 Function in Vivo* , 1996, The Journal of Biological Chemistry.

[30]  B. Vogelstein,et al.  A mammalian cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia , 1992, Cell.

[31]  Functional activation of p53 via phosphorylation following DNA damage by UV but not γ radiation , 1998 .

[32]  R Grosschedl,et al.  HMG domain proteins: architectural elements in the assembly of nucleoprotein structures. , 1994, Trends in genetics : TIG.

[33]  K. Isselbacher,et al.  Heterozygous germ line hCHK2 mutations in Li-Fraumeni syndrome. , 1999, Science.

[34]  A. Giaccia,et al.  The complexity of p53 modulation: emerging patterns from divergent signals. , 1998, Genes & development.

[35]  A. Levine,et al.  Non-p53 p53RE binding protein, a human transcription factor functionally analogous to P53. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[36]  T. Unger,et al.  Critical role for Ser20 of human p53 in the negative regulation of p53 by Mdm2 , 1999, The EMBO journal.

[37]  D. Lane,et al.  Regulation of the specific DNA binding function of p53 , 1992, Cell.

[38]  E. Olson,et al.  Cooperative Transcriptional Activation by Serum Response Factor and the High Mobility Group Protein SSRP1* , 1999, The Journal of Biological Chemistry.

[39]  S. Lowe,et al.  Oncogenic ras Provokes Premature Cell Senescence Associated with Accumulation of p53 and p16INK4a , 1997, Cell.

[40]  E. Appella,et al.  Phosphorylation of murine p53 at ser-18 regulates the p53 responses to DNA damage. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[41]  K. Sakaguchi,et al.  DNA damage activates p53 through a phosphorylation-acetylation cascade. , 1998, Genes & development.

[42]  Hua Lu,et al.  The N-Terminal Domain of p73 Interacts with the CH1 Domain of p300/CREB Binding Protein and Mediates Transcriptional Activation and Apoptosis , 2000, Molecular and Cellular Biology.

[43]  Yoichi Taya,et al.  DNA Damage-Induced Phosphorylation of p53 Alleviates Inhibition by MDM2 , 1997, Cell.

[44]  C. Harris,et al.  Nitric oxide-induced p53 accumulation and regulation of inducible nitric oxide synthase expression by wild-type p53. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[45]  T. Sugano,et al.  U.v.-induced nuclear accumulation of p53 is evoked through DNA damage of actively transcribed genes independent of the cell cycle. , 1994, Oncogene.

[46]  G. Wahl,et al.  A reversible, p53-dependent G0/G1 cell cycle arrest induced by ribonucleotide depletion in the absence of detectable DNA damage. , 1996, Genes & development.

[47]  A. Levine,et al.  Human TAFII31 protein is a transcriptional coactivator of the p53 protein. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[48]  T. Graeber,et al.  Hypoxia induces accumulation of p53 protein, but activation of a G1-phase checkpoint by low-oxygen conditions is independent of p53 status , 1994, Molecular and cellular biology.

[49]  B. Vogelstein,et al.  Participation of p53 protein in the cellular response to DNA damage. , 1991, Cancer research.

[50]  V. Rotter,et al.  Introduction: p53 – the first twenty years , 1999, Cellular and Molecular Life Sciences CMLS.

[51]  M. Yaffe,et al.  MAP kinase pathways activated by stress: the p38 MAPK pathway. , 2000, Critical care medicine.

[52]  D. Housman,et al.  Isolation and characterization of human cDNA clones encoding a high mobility group box protein that recognizes structural distortions to DNA caused by binding of the anticancer agent cisplatin. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[53]  Weiya Ma,et al.  p38 Kinase Mediates UV-induced Phosphorylation of p53 Protein at Serine 389* , 1999, The Journal of Biological Chemistry.

[54]  E. Egyházi.  Initiation inhibition and reinitiation of the synthesis of heterogeneous nuclear RNA in living cells , 1976, Nature.

[55]  F. Chen,et al.  Inhibition of RNA polymerase II as a trigger for the p53 response , 1999, Oncogene.

[56]  O. Issinger,et al.  CK2: a protein kinase in need of control. , 1999, Pharmacology & therapeutics.

[57]  D. Reinberg,et al.  Human general transcription factor IIH phosphorylates the C-terminal domain of RNA polymerase II , 1992, Nature.

[58]  G. Orphanides,et al.  The chromatin-specific transcription elongation factor FACT comprises human SPT16 and SSRP1 proteins , 1999, Nature.