title: Non-canonical CRL4A/4B(CDT2) interacts with RAD18 to modulate post replication repair and cell survival.
abstract: The Cullin-4(CDT2) E3 ubiquitin ligase plays an essential role in DNA replication origin licensing directing degradation of several licensing factors at the G1/S transition in order to prevent DNA re-replication. Recently a RAD18-independent role of Cullin-4(CDT2) in PCNA monoubiquitylation has been proposed. In an effort to better understand the function of Cullin-4(CDT2) E3 ubiquitin ligase in mammalian Post-Replication Repair during an unperturbed S-phase, we show that down-regulation of Cullin-4(CDT2) leads to two distinguishable independent phenotypes in human cells that unveil at least two independent roles of Cullin-4(CDT2) in S-phase. Apart from the re-replication preventing activity, we identified a non-canonical Cullin-4(CDT2) complex, containing both CUL4A and CUL4B, associated to the COP9 signalosome, that controls a RAD18-dependent damage avoidance pathway essential during an unperturbed S-phase. Indeed, we show that the non-canonical Cullin-4A/4B(CDT2) complex binds to RAD18 and it is required to modulate RAD18 protein levels onto chromatin and the consequent dynamics of PCNA monoubiquitylation during a normal S-phase. This function prevents replication stress, ATR hyper-signaling and, ultimately, apoptosis. A very similar PRR regulatory mechanism has been recently described for Spartan. Our findings uncover a finely regulated process in mammalian cells involving Post-Replication Repair factors, COP9 signalosome and a non-canonical Cullin4-based E3 ligase which is essential to tolerate spontaneous damage and for cell survival during physiological DNA replication.
authors.affiliation: Dipartimento di Bioscienze, Università degli Studi di Milano, Milano, Italy.
authors.forename: Sarah
authors.initials: S
authors.lastname: Sertic
authors.forename: Claudio
authors.initials: C
authors.lastname: Evolvi
authors.forename: Emanuela
authors.initials: E
authors.lastname: Tumini
authors.forename: Paolo
authors.initials: P
authors.lastname: Plevani
authors.forename: Marco
authors.initials: M
authors.lastname: Muzi-Falconi
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Rotondo
mesh_terms: D017209:Apoptosis / Q000235:genetics / Q000502:physiology*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics / Q000502:physiology; D004261:DNA Replication / Q000235:genetics / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; C429530:DTL protein, human; D009687:Nuclear Proteins; C485954:RAD18 protein, human; D044767:Ubiquitin-Protein Ligases
references.citation: Microbiol Mol Biol Rev. 2009 Mar;73(1):134-54
references.pmid: 19258535
references.citation: Nat Rev Mol Cell Biol. 2010 Mar;11(3):220-8
references.pmid: 20177397
references.citation: Cancer Cell. 2010 Oct 19;18(4):329-40
references.pmid: 20951943
references.citation: Cell Cycle. 2006 Aug;5(15):1719-29
references.pmid: 16861890
references.citation: Genes Dev. 2007 Nov 15;21(22):2908-22
references.pmid: 18006686
references.citation: Mol Cell. 2006 Sep 1;23(5):709-21
references.pmid: 16949367
references.citation: DNA Repair (Amst). 2010 Mar 2;9(3):257-67
references.pmid: 20096653
references.citation: Genes Dev. 2006 Nov 15;20(22):3117-29
references.pmid: 17085480
references.citation: Nat Cell Biol. 2004 Oct;6(10):1003-9
references.pmid: 15448697
references.citation: Mutat Res. 2001 Aug 9;486(3):167-84
references.pmid: 11459630
references.citation: Cell. 2011 Mar 4;144(5):646-74
references.pmid: 21376230
references.citation: Genes Cells. 2009 Nov;14(11):1209-25
references.pmid: 19849719
references.citation: Mol Cell Biol. 2006 Nov;26(21):7977-90
references.pmid: 16940174
references.citation: EMBO J. 2004 Oct 1;23(19):3886-96
references.pmid: 15359278
references.citation: Development. 2008 Jun;135(11):2013-22
references.pmid: 18434413
references.citation: Nat Cell Biol. 2010 Nov;12(11):1086-93
references.pmid: 20953199
references.citation: Mol Cell. 2009 Sep 11;35(5):586-97
references.pmid: 19748355
references.citation: Science. 2003 Jun 6;300(5625):1542-8
references.pmid: 12791985
references.citation: J Cell Biol. 2011 Jan 10;192(1):43-54
references.pmid: 21220508
references.citation: J Biol Chem. 2008 Oct 24;283(43):29045-52
references.pmid: 18703516
references.citation: Cell Cycle. 2006 Aug;5(15):1675-80
references.pmid: 16861906
references.citation: Mol Cells. 2008 Jul 31;26(1):5-11
references.pmid: 18525240
references.citation: Genes Dev. 2005 May 1;19(9):1040-52
references.pmid: 15833913
references.citation: Int J Cancer. 2006 Dec 15;119(12):2768-74
references.pmid: 17036332
references.citation: Genes Dev. 2008 Sep 15;22(18):2496-506
references.pmid: 18794347
references.citation: Cell. 2007 Jun 15;129(6):1165-76
references.pmid: 17574027
references.citation: Front Oncol. 2012 Apr 09;2:29
references.pmid: 22655267
references.citation: Curr Protein Pept Sci. 2004 Jun;5(3):185-9
references.pmid: 15180523
references.citation: Curr Opin Struct Biol. 2010 Dec;20(6):714-21
references.pmid: 20880695
references.citation: Trends Biochem Sci. 2009 Nov;34(11):562-70
references.pmid: 19818632
references.citation: Mol Cell. 2008 Dec 26;32(6):757-66
references.pmid: 19111656
references.citation: Cancer Res. 2007 Nov 15;67(22):10899-909
references.pmid: 18006835
references.citation: Mol Biol Cell. 2002 Jun;13(6):1977-2000
references.pmid: 12058064
references.citation: Mol Cell. 2012 Jun 8;46(5):625-35
references.pmid: 22681887
references.citation: Cell. 2003 May 2;113(3):357-67
references.pmid: 12732143
references.citation: Mol Cell. 2010 Oct 8;40(1):9-21
references.pmid: 20932471
references.citation: Nat Cell Biol. 2006 Apr;8(4):339-47
references.pmid: 16531995
references.citation: Cell Cycle. 2011 Jan 15;10(2):241-9
references.pmid: 21212733
references.citation: Nat Cell Biol. 2003 Nov;5(11):1008-15
references.pmid: 14578910
references.citation: Mol Cell. 2010 Nov 12;40(3):364-76
references.pmid: 21035370
references.citation: Cell Signal. 2007 May;19(5):1071-80
references.pmid: 17254749
references.citation: Mol Cell. 2010 Oct 8;40(1):22-33
references.pmid: 20932472
references.citation: Hum Mol Genet. 2010 Apr 1;19(7):1324-34
references.pmid: 20064923
references.citation: EMBO J. 2006 Mar 8;25(5):1126-36
references.pmid: 16482215
references.citation: Mol Cell. 2011 Apr 22;42(2):237-49
references.pmid: 21396873
references.citation: Am J Pathol. 2004 Oct;165(4):1351-65
references.pmid: 15466399
references.citation: DNA Repair (Amst). 2010 Dec 10;9(12):1241-8
references.pmid: 20971043
references.citation: Cancer Res. 1999 Sep 1;59(17):4375-82
references.pmid: 10485486
references.citation: Mol Biol Cell. 2007 Mar;18(3):899-909
references.pmid: 17192413
references.citation: DNA Repair (Amst). 2011 Feb 7;10(2):149-58
references.pmid: 21109493
references.citation: Subcell Biochem. 2010;54:57-68
references.pmid: 21222273
references.citation: Cell Res. 2008 Jan;18(1):162-73
references.pmid: 18157158
references.citation: J Mol Cell Biol. 2011 Feb;3(1):13-22
references.pmid: 21278447
references.citation: Mol Cell. 2010 Jan 15;37(1):143-9
references.pmid: 20129063