title: RAD6 regulates the dosage of p53 by a combination of transcriptional and posttranscriptional mechanisms.
abstract: Maintaining an appropriate cellular concentration of p53 is critical for cell survival and normal development in various organisms. In this study, we provide evidence that the human E2 ubiquitin-conjugating enzyme RAD6 plays a critical role in regulating p53 protein levels under both normal and stress conditions. Knockdown and overexpression of RAD6 affected p53 turnover and transcription. We showed that RAD6 can form a ternary complex with MDM2 and p53 that contributes to the degradation of p53. Chromatin immunoprecipitation (ChIP) analysis showed that RAD6 also binds to the promoter and coding regions of the p53 gene and modulates the levels of H3K4 and K79 methylation on local chromatin. When the cells were exposed to stress stimuli, the RAD6-MDM2-p53 ternary complex was disrupted; RAD6 was then recruited to the chromatin of the p53 gene, resulting in an increase in histone methylation and p53 transcription. Further studies showed that stress-induced p53 transcriptional activation, cell apoptosis, and disrupted cell cycle progression are all RAD6 dependent. Overall, this work demonstrates that RAD6 regulates p53 levels in a "yin-yang" manner through a combination of two distinct mechanisms in mammalian cells.
authors.affiliation: Institute of Epigenetics and Cancer Research, School of Medicine, Tsinghua University, Beijing, China.
authors.forename: Su
authors.initials: S
authors.lastname: Chen
authors.forename: Da-Liang
authors.initials: DL
authors.lastname: Wang
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Lei
authors.initials: L
authors.lastname: Zhao
authors.forename: Fang-Lin
authors.initials: FL
authors.lastname: Sun
mesh_terms: D002843:Chromatin / Q000235:genetics / Q000378:metabolism; D047369:Chromatin Immunoprecipitation; D005786:Gene Expression Regulation; D016158:Genes, p53; D006367:HeLa Cells; D006657:Histones / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008745:Methylation; D011401:Promoter Regions, Genetic; D051736:Proto-Oncogene Proteins c-mdm2 / Q000378:metabolism; D015533:Transcriptional Activation; D016159:Tumor Suppressor Protein p53 / Q000235:genetics* / Q000378:metabolism; D044763:Ubiquitin-Conjugating Enzymes / Q000235:genetics / Q000378:metabolism*
chemical_list: D002843:Chromatin; D006657:Histones; D016159:Tumor Suppressor Protein p53; C471545:UBE2A protein, human; C471546:UBE2B protein, human; D044763:Ubiquitin-Conjugating Enzymes; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2
references.citation: J Biol Chem. 2011 Mar 18;286(11):9020-30
references.pmid: 21205821
references.citation: Mol Cell. 2005 Dec 9;20(5):759-70
references.pmid: 16337599
references.citation: J Biol Chem. 2002 Sep 20;277(38):34655-7
references.pmid: 12167634
references.citation: Science. 1989 Mar 24;243(4898):1576-83
references.pmid: 2538923
references.citation: Mol Cell. 2005 Nov 23;20(4):601-11
references.pmid: 16307923
references.citation: Genes Dev. 1993 Feb;7(2):250-61
references.pmid: 8436296
references.citation: Nature. 2002 Jul 4;418(6893):104-8
references.pmid: 12077605
references.citation: Mol Cell. 2007 Jan 12;25(1):15-30
references.pmid: 17218268
references.citation: Cell. 2010 Feb 5;140(3):384-96
references.pmid: 20096447
references.citation: Curr Opin Genet Dev. 2005 Feb;15(1):27-33
references.pmid: 15661530
references.citation: Cell. 1994 Oct 7;79(1):13-21
references.pmid: 7923371
references.citation: Cell. 1997 Feb 7;88(3):323-31
references.pmid: 9039259
references.citation: Adv Genet. 1982;21:173-254
references.pmid: 7036692
references.citation: Mol Cell Biol. 2003 Apr;23(7):2463-75
references.pmid: 12640129
references.citation: Proc Natl Acad Sci U S A. 2002 Jan 8;99(1):90-4
references.pmid: 11752412
references.citation: Nature. 1987 Sep 10-16;329(6135):131-4
references.pmid: 3306404
references.citation: Nucleic Acids Res. 2010 Oct;38(19):6544-54
references.pmid: 20542919
references.citation: Nature. 2005 Jul 21;436(7049):428-33
references.pmid: 15931174
references.citation: Proc Natl Acad Sci U S A. 1990 Apr;87(7):2695-9
references.pmid: 2157209
references.citation: Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7351-5
references.pmid: 1651502
references.citation: J Biol Chem. 1997 Sep 12;272(37):23360-5
references.pmid: 9287349
references.citation: Science. 2003 Dec 12;302(5652):1972-5
references.pmid: 14671306
references.citation: Mol Cell Biol. 2004 Jun;24(11):4710-9
references.pmid: 15143166
references.citation: Proc Natl Acad Sci U S A. 2001 Nov 6;98(23):12902-7
references.pmid: 11687631
references.citation: Mol Cell. 2005 Nov 23;20(4):589-99
references.pmid: 16307922
references.citation: J Biol Chem. 2003 Sep 12;278(37):34739-42
references.pmid: 12876294
references.citation: FEBS Lett. 1997 Dec 22;420(1):25-7
references.pmid: 9450543
references.citation: Nature. 2002 Sep 12;419(6903):135-41
references.pmid: 12226657
references.citation: EMBO J. 2004 Nov 24;23(23):4583-94
references.pmid: 15526030
references.citation: Proc Natl Acad Sci U S A. 1991 May 1;88(9):3832-6
references.pmid: 1902572
references.citation: Proc Natl Acad Sci U S A. 2009 Jul 14;106(28):11612-6
references.pmid: 19556538
references.citation: Cell. 2009 May 1;137(3):459-71
references.pmid: 19410543
references.citation: Dev Biol. 1996 Jan 10;173(1):119-32
references.pmid: 8575614
references.citation: Mol Cells. 2008 Jul 31;26(1):5-11
references.pmid: 18525240
references.citation: Cell. 2007 Dec 14;131(6):1084-96
references.pmid: 18083099
references.citation: Mol Cell Biol. 2003 Feb;23(4):1151-62
references.pmid: 12556476
references.citation: Genes Dev. 2008 Oct 1;22(19):2664-76
references.pmid: 18832071
references.citation: Trends Cell Biol. 2010 Mar;20(3):170-5
references.pmid: 20061153
references.citation: EMBO J. 2001 Dec 17;20(24):7137-48
references.pmid: 11742990
references.citation: Genes Dev. 1994 Apr 1;8(7):811-20
references.pmid: 7926769
references.citation: Mol Cell. 2006 Feb 3;21(3):307-15
references.pmid: 16455486
references.citation: Annu Rev Biochem. 2006;75:243-69
references.pmid: 16756492
references.citation: Semin Cancer Biol. 2003 Feb;13(1):49-58
references.pmid: 12507556
references.citation: Curr Biol. 1992 Nov;2(11):581-3
references.pmid: 15336029
references.citation: Cell. 1989 Jun 30;57(7):1083-93
references.pmid: 2525423
references.citation: Curr Opin Cell Biol. 1995 Apr;7(2):215-23
references.pmid: 7612274
references.citation: Mol Biol Cell. 1997 Dec;8(12):2421-36
references.pmid: 9398665
references.citation: Cell. 1996 Sep 6;86(5):799-810
references.pmid: 8797826
references.citation: Mol Cell Biol. 2004 Jun;24(12):5485-95
references.pmid: 15169909
references.citation: Nature. 1997 May 15;387(6630):299-303
references.pmid: 9153396
references.citation: Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8865-9
references.pmid: 1717990
references.citation: Cell. 2006 May 19;125(4):703-17
references.pmid: 16713563
references.citation: J Cell Sci. 2007 Jun 1;120(Pt 11):1841-51
references.pmid: 17488778
references.citation: Proc Natl Acad Sci U S A. 2004 Aug 3;101(31):11380-5
references.pmid: 15280549
references.citation: Oncogene. 2004 Apr 15;23(17):3097-106
references.pmid: 14981545
references.citation: J Biol Chem. 2002 Aug 9;277(32):28368-71
references.pmid: 12070136
references.citation: Nat Cell Biol. 2009 May;11(5):580-91
references.pmid: 19377469
references.citation: Nature. 2007 May 24;447(7143):407-12
references.pmid: 17522673
references.citation: J Biol Chem. 2002 Mar 29;277(13):10753-5
references.pmid: 11805083
references.citation: Genes Dev. 2006 Jul 15;20(14):1959-73
references.pmid: 16816001
references.citation: Nature. 1997 May 15;387(6630):296-9
references.pmid: 9153395
references.citation: Mol Cell. 2003 Jan;11(1):267-74
references.pmid: 12535539
references.citation: Cancer Res. 1991 Aug 15;51(16):4279-86
references.pmid: 1868448
references.citation: Nat Rev Cancer. 2002 Aug;2(8):594-604
references.pmid: 12154352
references.citation: Cell. 2009 May 15;137(4):609-22
references.pmid: 19450511
references.citation: Science. 1989 Apr 14;244(4901):217-21
references.pmid: 2649981
references.citation: EMBO J. 1991 Aug;10(8):2187-93
references.pmid: 2065660
references.citation: J Biol Chem. 2009 Jul 31;284(31):20582-92
references.pmid: 19531475
references.citation: Mol Cell Biol. 1997 Aug;17(8):4536-43
references.pmid: 9234711