Involvement of PIAS1 in the sumoylation of tumor suppressor p53.

Sumoylation of p53 by the ubiquitin-like protein, SUMO-1/sentrin/PIC1, has been shown to stimulate its transcriptional activation activity. The SUMO E3 ligase, a key enzyme in the recognition of substrates to be sumoylated, has not yet been identified. We isolated PIAS1 (protein inhibitor of activated STAT1) as a SUMO-1 binding protein by yeast two-hybrid screening. In addition, PIAS1 bound p53 and Ubc9, the E2 for SUMO. PIAS1 that was mutated in the RING finger-like domain bound p53 and SUMO-1, but not Ubc9. PIAS1 catalyzed the sumoylation of p53 both in U2OS cells and in vitro in a domain-dependent manner. These data suggest that PIAS1 functions as a SUMO ligase, or possibly as a tightly bound regulator of it, toward p53.

[1]  F. Melchior,et al.  Modification of Ran GTPase-activating Protein by the Small Ubiquitin-related Modifier SUMO-1 Requires Ubc9, an E2-type Ubiquitin-conjugating Enzyme Homologue* , 1998, The Journal of Biological Chemistry.

[2]  S. Jentsch,et al.  Ubiquitin and proteasomes: Sumo, ubiquitin's mysterious cousin , 2001, Nature Reviews Molecular Cell Biology.

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

[4]  R. Baron,et al.  Ligand-induced Ubiquitination of the Epidermal Growth Factor Receptor Involves the Interaction of the c-Cbl RING Finger and UbcH7* , 1999, The Journal of Biological Chemistry.

[5]  D. Chang,et al.  Inhibition of Stat1-mediated gene activation by PIAS1. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Wei Gu,et al.  Activation of p53 Sequence-Specific DNA Binding by Acetylation of the p53 C-Terminal Domain , 1997, Cell.

[7]  R. Honda,et al.  In vitro SUMO-1 modification requires two enzymatic steps, E1 and E2. , 1999, Biochemical and biophysical research communications.

[8]  T. Hunter,et al.  The tyrosine kinase negative regulator c-Cbl as a RING-type, E2-dependent ubiquitin-protein ligase. , 1999, Science.

[9]  A. Ciechanover,et al.  The ubiquitin system. , 1998, Annual review of biochemistry.

[10]  M. Dasso,et al.  RanBP2 associates with Ubc9p and a modified form of RanGAP1. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Fred Schaper,et al.  The zinc finger protein Gfi‐1 can enhance STAT3 signaling by interacting with the STAT3 inhibitor PIAS3 , 2000, The EMBO journal.

[12]  S. Berger,et al.  p53 Sites Acetylated In Vitro by PCAF and p300 Are Acetylated In Vivo in Response to DNA Damage , 1999, Molecular and Cellular Biology.

[13]  E. Yeh,et al.  Molecular cloning and characterization of human AOS1 and UBA2, components of the sentrin‐activating enzyme complex , 1999, FEBS letters.

[14]  K. Shuai,et al.  Modulation of STAT signaling by STAT-interacting proteins , 2000, Oncogene.

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

[16]  Wei Zhu,et al.  Arginine Methylation of STAT1 Modulates IFNα/β-Induced Transcription , 2001, Cell.

[17]  T. Sternsdorf,et al.  Evidence for Covalent Modification of the Nuclear Dot–associated Proteins PML and Sp100 by PIC1/SUMO-1 , 1997, The Journal of cell biology.

[18]  J. ten Hoeve,et al.  A transcriptional corepressor of Stat1 with an essential LXXLL signature motif , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[19]  R. Hay,et al.  SUMO-1 modification of IkappaBalpha inhibits NF-kappaB activation. , 1998, Molecular cell.

[20]  S. Fang,et al.  RING fingers mediate ubiquitin-conjugating enzyme (E2)-dependent ubiquitination. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[21]  K. Shuai,et al.  Distinct roles of the NH2- and COOH-terminal domains of the protein inhibitor of activated signal transducer and activator of transcription (STAT) 1 (PIAS1) in cytokine-induced PIAS1-Stat1 interaction. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[22]  R. Honda,et al.  Activity of MDM2, a ubiquitin ligase, toward p53 or itself is dependent on the RING finger domain of the ligase , 2000, Oncogene.

[23]  R. Hay,et al.  Identification of the Enzyme Required for Activation of the Small Ubiquitin-like Protein SUMO-1* , 1999, The Journal of Biological Chemistry.

[24]  P Jay,et al.  Specific inhibition of Stat3 signal transduction by PIAS3. , 1997, Science.

[25]  A. Hengstermann,et al.  Activation of p53 by conjugation to the ubiquitin‐like protein SUMO‐1 , 1999, The EMBO journal.

[26]  P. Freemont,et al.  SUMO-1 modification of the acute promyelocytic leukaemia protein PML: implications for nuclear localisation. , 1999, Journal of cell science.

[27]  A. Ciechanover,et al.  The ubiquitin-proteasome pathway: the complexity and myriad functions of proteins death. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[28]  O. Silvennoinen,et al.  ARIP3 (androgen receptor-interacting protein 3) and other PIAS (protein inhibitor of activated STAT) proteins differ in their ability to modulate steroid receptor-dependent transcriptional activation. , 2000, Molecular endocrinology.

[29]  A. Dejean,et al.  Conjugation with the ubiquitin‐related modifier SUMO‐1 regulates the partitioning of PML within the nucleus , 1998, The EMBO journal.

[30]  L. Bracco,et al.  MBP1: a novel mutant p53-specific protein partner with oncogenic properties , 1999, Oncogene.

[31]  Hirofumi Tanaka,et al.  Oncoprotein MDM2 is a ubiquitin ligase E3 for tumor suppressor p53 , 1997, FEBS letters.

[32]  R. Hay,et al.  SUMO‐1 modification activates the transcriptional response of p53 , 1999, The EMBO journal.