title: DCNL1 functions as a substrate sensor and activator of cullin 2-RING ligase.
abstract: Substrate engagement by F-box proteins promotes NEDD8 modification of cullins, which is necessary for the activation of cullin-RING E3 ubiquitin ligases (CRLs). However, the mechanism by which substrate recruitment triggers cullin neddylation remains unclear. Here, we identify DCNL1 (defective in cullin neddylation 1-like 1) as a component of CRL2 called ECV (elongins BC/CUL2/VHL) and show that molecular suppression of DCNL1 attenuates CUL2 neddylation. DCNL1 via its DAD patch binds to CUL2 but is also able to bind VHL independent of CUL2 and the DAD patch. The engagement of the substrate hypoxia-inducible factor 1α (HIF1α) to the substrate receptor VHL increases DCNL1 binding to VHL as well as to CUL2. Notably, an engineered mutant form of HIF1α that associates with CUL2, but not DCNL1, fails to trigger CUL2 neddylation and retains ECV in an inactive state. These findings support a model in which substrate engagement prompts DCNL1 recruitment that facilitates the initiation of CUL2 neddylation and define DCNL1 as a "substrate sensor switch" for ECV activation.
authors: Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, Ontario, Canada.
authors: Pardeep
authors: P
authors: Heir
authors: Roxana I
authors: RI
authors: Sufan
authors: Samantha N
authors: SN
authors: Greer
authors: Betty P
authors: BP
authors: Poon
authors: Jeffrey E
authors: JE
authors: Lee
authors: Michael
authors: M
authors: Ohh
mesh_terms: D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D044842:Cullin Proteins / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D009154:Mutation; D011485:Protein Binding; D011506:Proteins; D011518:Proto-Oncogene Proteins; D034622:RNA Interference; D034741:RNA, Small Interfering; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000378:metabolism*
chemical_list: C105153:CUL2 protein, human; D018797:Cell Cycle Proteins; D044842:Cullin Proteins; C408631:DCUN1D1 protein, human; C497442:HIF1A protein, human; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D047908:Intracellular Signaling Peptides and Proteins; D011506:Proteins; D011518:Proto-Oncogene Proteins; D034741:RNA, Small Interfering; D044767:Ubiquitin-Protein Ligases; D051794:Von Hippel-Lindau Tumor Suppressor Protein; C497446:VHL protein, human
references: Cell. 2010 Dec 10;143(6):951-65
references: 21145461
references: J Biol Chem. 1999 Dec 10;274(50):36025-9
references: 10585493
references: Biochem Biophys Res Commun. 1999 Apr 2;257(1):100-5
references: 10092517
references: Science. 2002 Jun 7;296(5574):1886-9
references: 12004076
references: Mol Cell. 2008 May 23;30(4):393-402
references: 18498744
references: Nat Cell Biol. 2000 Jul;2(7):423-7
references: 10878807
references: Science. 2001 Apr 20;292(5516):464-8
references: 11292862
references: EMBO J. 1998 Apr 15;17(8):2208-14
references: 9545234
references: Curr Opin Struct Biol. 2010 Dec;20(6):714-21
references: 20880695
references: N Engl J Med. 2011 Aug 11;365(6):537-47
references: 21830968
references: Science. 2001 Nov 9;294(5545):1337-40
references: 11598268
references: Mol Cell. 2008 Oct 10;32(1):21-31
references: 18851830
references: Cell. 2008 Sep 19;134(6):995-1006
references: 18805092
references: Science. 1999 Apr 16;284(5413):455-61
references: 10205047
references: Proc Natl Acad Sci U S A. 1999 May 11;96(10):5510-5
references: 10318914
references: J Biol Chem. 2007 Jun 8;282(23):17032-40
references: 17439941
references: Proc Natl Acad Sci U S A. 2009 Jul 28;106(30):12365-70
references: 19617556
references: J Biol Chem. 2003 Mar 28;278(13):11032-40
references: 12538644
references: Cancer Cell. 2011 Feb 15;19(2):168-76
references: 21316600
references: Mol Cell Biol. 1998 Feb;18(2):732-41
references: 9447969
references: Neoplasia. 2006 Nov;8(11):956-63
references: 17132228
references: Genes Dev. 1998 Aug 1;12(15):2263-8
references: 9694792
references: Proc Natl Acad Sci U S A. 2006 Aug 1;103(31):11515-20
references: 16861300
references: Nat Struct Mol Biol. 2011 Jul 17;18(8):947-9
references: 21765416
references: Mol Cell. 2010 Sep 10;39(5):784-96
references: 20832729
references: Nat Rev Mol Cell Biol. 2009 May;10(5):319-31
references: 19352404
references: Cell. 2001 Oct 5;107(1):43-54
references: 11595184
references: J Biol Chem. 2001 Apr 20;276(16):12530-8
references: 11278610
references: Mol Cell. 2008 Jan 18;29(1):23-35
references: 18206966
references: Nature. 2005 Jun 30;435(7046):1257-61
references: 15988528
references: EMBO Rep. 2002 Feb;3(2):177-82
references: 11818338
references: Science. 2001 Apr 20;292(5516):468-72
references: 11292861
references: Mol Cell Biol. 2004 Apr;24(8):3251-61
references: 15060148
references: Genes Dev. 2004 Dec 15;18(24):3055-65
references: 15601820
references: Oncogene. 1999 Nov 18;18(48):6829-34
references: 10597293
references: J Biol Chem. 2008 Nov 28;283(48):33211-20
references: 18826954
references: Mol Cell Biol. 2000 Apr;20(7):2326-33
references: 10713156
references: Nat Rev Mol Cell Biol. 2005 Jan;6(1):9-20
references: 15688063
references: Nature. 2002 Jun 27;417(6892):975-8
references: 12050673
references: PLoS One. 2011;6(9):e25358
references: 21980433
references: EMBO J. 2001 Aug 1;20(15):4003-12
references: 11483504