TIP30 has an intrinsic kinase activity required for up‐regulation of a subset of apoptotic genes

CC3 is a metastasis suppressor that inhibits metastasis of the variant small cell lung carcinoma (v‐SCLC) by predisposing cells to apoptosis. The same protein was also reported as a cellular cofactor, TIP30, which stimulates HIV‐1 Tat‐activated transcription by interacting with both Tat and RNA polymerase II. We report here that TIP30/CC3 is a novel serine/threonine kinase. It phosphorylates the heptapeptide repeats of the C‐terminal domain (CTD) of the largest RNA polymerase II subunit in a Tat‐dependent manner. Amino acid substitutions in the putative ATP binding motif that abolish the TIP30 kinase activity also inhibit the ability of TIP30 to enhance Tat‐activated transcription or to sensitize NIH 3T3 and v‐SCLC cells to apoptosis. Furthermore, ectopic expression of TIP30/CC3 in v‐SCLC cells induces expression of a number of genes that include the apoptosis‐related genes Bad and Siva, as well as metastasis suppressor NM23‐H2. These data demonstrate a molecular mechanism for TIP30/CC3 function and suggest a novel pathway for regulating apoptosis.

[1]  K. Jeang,et al.  Activation of Integrated Provirus Requires Histone Acetyltransferase , 1998, The Journal of Biological Chemistry.

[2]  S. J. Flint,et al.  Human c-myc transcription factor PuF identified as nm23-H2 nucleoside diphosphate kinase, a candidate suppressor of tumor metastasis. , 1993, Science.

[3]  K. Kinzler,et al.  A model for p53-induced apoptosis , 1997, Nature.

[4]  M. Garber,et al.  The interaction between HIV-1 Tat and human cyclin T1 requires zinc and a critical cysteine residue that is not conserved in the murine CycT1 protein. , 1998, Genes & development.

[5]  H. Okamoto,et al.  Trans-activation by human immunodeficiency virus Tat protein requires the C-terminal domain of RNA polymerase II. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[6]  D. Price,et al.  Purification of P-TEFb, a Transcription Factor Required for the Transition into Productive Elongation (*) , 1995, The Journal of Biological Chemistry.

[7]  Z. Ao,et al.  CD27, a member of the tumor necrosis factor receptor family, induces apoptosis and binds to Siva, a proapoptotic protein. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[8]  R. Roeder,et al.  Regulation of TFIIH ATPase and kinase activities by TFIIE during active initiation complex formation , 1994, Nature.

[9]  A. Thor,et al.  Systemic Gene Delivery Expands the Repertoire of Effective Antiangiogenic Agents* , 1999, The Journal of Biological Chemistry.

[10]  N. Hay,et al.  Myc-mediated apoptosis requires wild-type p53 in a manner independent of cell cycle arrest and the ability of p53 to induce p21waf1/cip1. , 1994, Genes & development.

[11]  H. Okamoto,et al.  The HIV transactivator TAT binds to the CDK-activating kinase and activates the phosphorylation of the carboxy-terminal domain of RNA polymerase II. , 1997, Genes & development.

[12]  Gerard I. Evan,et al.  Induction of apoptosis in fibroblasts by c-myc protein , 1992, Cell.

[13]  J. Greenblatt,et al.  A cofactor, TIP30, specifically enhances HIV-1 Tat-activated transcription. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Roeder,et al.  Expression and purification of general transcription factors by FLAG epitope-tagging and peptide elution. , 1993, Peptide research.

[15]  D. Bossemeyer,et al.  The glycine-rich sequence of protein kinases: a multifunctional element. , 1994, Trends in biochemical sciences.

[16]  R. Young,et al.  A kinase–cyclin pair in the RNA polymerase II holoenzyme , 1995, Nature.

[17]  H. Hermeking,et al.  Mediation of c-Myc-induced apoptosis by p53. , 1994, Science.

[18]  R. Roeder,et al.  Enhanced processivity of RNA polymerase II triggered by Tat-induced phosphorylation of its carboxy-terminal domain , 1996, Nature.

[19]  M. Baker,et al.  TIP30, a cofactor for HIV-1 Tat-activated transcription, is homologous to short-chain dehydrogenases/reductases , 1999, Current Biology.

[20]  M. Dahmus.  Phosphorylation of the C-terminal domain of RNA polymerase II. , 1995, Biochimica et biophysica acta.

[21]  H. Galjaard.  Biochemical analysis of single cultured cells , 1980 .

[22]  Ping Wei,et al.  A Novel CDK9-Associated C-Type Cyclin Interacts Directly with HIV-1 Tat and Mediates Its High-Affinity, Loop-Specific Binding to TAR RNA , 1998, Cell.

[23]  C. Rudin,et al.  Signal transduction pathways that regulate cell survival and cell death , 1998, Oncogene.

[24]  G. Evan,et al.  Suppression of c-Myc-induced apoptosis by Ras signalling through PI(3)K and PKB , 1997, Nature.

[25]  J. Yokota,et al.  Induction of apoptosis but not G1 arrest by expression of the wild-type p53 gene in small cell lung carcinoma. , 1996, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[26]  J. Milton,et al.  Identification of multiple cyclin subunits of human P-TEFb. , 1998, Genes & development.

[27]  John Calvin Reed,et al.  Tumor suppressor p53 is a direct transcriptional activator of the human bax gene , 1995, Cell.

[28]  D. Thorley-Lawson,et al.  A novel form of Epstein-Barr virus latency in normal B cells in vivo , 1995, Cell.

[29]  M. Giacca,et al.  HIV-1 tat transactivator recruits p300 and CREB-binding protein histone acetyltransferases to the viral promoter. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[30]  E. Shtivelman.  A link between metastasis and resistance to apoptosis of variant small cell lung carcinoma , 1997, Oncogene.

[31]  Jean Y. J. Wang,et al.  Tyrosine Phosphorylation of RNA Polymerase II Carboxyl-terminal Domain by the Abl-related Gene Product* , 1997, The Journal of Biological Chemistry.

[32]  A. Bassuk,et al.  Physical interactions between Ets and NF-kappaB/NFAT proteins play an important role in their cooperative activation of the human immunodeficiency virus enhancer in T cells , 1997, Journal of virology.

[33]  C. Anderson,et al.  DNA binding provides a signal for phosphorylation of the RNA polymerase II heptapeptide repeats. , 1992, Genes & development.

[34]  D. Price,et al.  Control of RNA Polymerase II Elongation Potential by a Novel Carboxyl-terminal Domain Kinase* , 1996, The Journal of Biological Chemistry.

[35]  G. Nabel,et al.  Interaction of Human Immunodeficiency Virus Type 1 Tat with the Transcriptional Coactivators p300 and CREB Binding Protein , 1998, Journal of Virology.

[36]  J. Greenblatt,et al.  Three functional classes of transcriptional activation domain , 1996, Molecular and cellular biology.

[37]  S. Korsmeyer,et al.  Bad, a heterodimeric partner for Bcl-xL and Bcl-2, displaces bax and promotes cell death , 1995, Cell.

[38]  J C Reed,et al.  Mitochondria and apoptosis. , 1998, Science.

[39]  K. Jeang,et al.  Requirements for RNA Polymerase II Carboxyl-terminal Domain for Activated Transcription of Human Retroviruses Human T-Cell Lymphotropic Virus I and HIV-1* , 1996, The Journal of Biological Chemistry.

[40]  C. Marshall,et al.  Hypertrophic Agonists Stimulate the Activities of the Protein Kinases c-Raf and A-Raf in Cultured Ventricular Myocytes (*) , 1995, The Journal of Biological Chemistry.

[41]  D. Hazuda,et al.  P-TEFb kinase is required for HIV Tat transcriptional activation in vivo and in vitro. , 1997, Genes & development.

[42]  B. Cullen,et al.  Recruitment of a protein complex containing Tat and cyclin T1 to TAR governs the species specificity of HIV‐1 Tat , 1998, The EMBO journal.

[43]  T. Hunter,et al.  The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. , 1988, Science.

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

[45]  M. Mathews,et al.  Transcription elongation factor P-TEFb is required for HIV-1 tat transactivation in vitro. , 1997, Genes & development.

[46]  S. Korsmeyer,et al.  Cell Death in Development , 1999, Cell.