title: Centrosomes at M phase act as a scaffold for the accumulation of intracellular ubiquitinated proteins.
abstract: Centrosome size varies considerably during the cell cycle; it is greatest during metaphase, partly because of pericentriolar matrix recruitment and an increase in microtubule-organizing activity. However, the mechanism of centrosome maturation during M phase is poorly defined. In the present study, we identified and quantified centrosomal proteins during S and M phases using stable isotope labeling by amino acids in cell culture (SILAC) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). We identified 991 proteins, of which 310 and 325 proteins were upregulated during S and M phases, respectively. Ubiquitinated proteins containing K48- and K63-linked polyubiquitin chains accumulated in the centrosomes during M phase, although 26S proteasome activity in the centrosomes did not markedly differ between S and M phases. Conversely, cytoplasmic dynein, which transports ubiquitinated proteins to the centrosomes, increased 2-fold in the centrosomes during M phase relative to S phase. Furthermore, PYR-41, a ubiquitin E1 inhibitor, reduced centrosome size during metaphase, causing increased aneuploidy. RNA interference suppression of Ecm29, which inhibits proteasome activity, decreased the accumulation of ubiquitinated proteins in the centrosomes. These results show that accumulation of ubiquitinated proteins promotes centrosome maturation during M phase and further suggest a novel function of centrosomes as a scaffold temporarily gathering intracellular ubiquitinated proteins.
authors.affiliation: Department of Molecular Genetics; Medical Research Institute; Tokyo Medical and Dental University (TMDU); Bunkyo-ku, Tokyo, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Kimura
authors.affiliation: Department of Molecular Genetics; Medical Research Institute; Tokyo Medical and Dental University (TMDU); Bunkyo-ku, Tokyo, Japan; Department of Genetic Diagnosis; The Cancer Institute; Japanese Foundation for Cancer Research; Koto-ku, Tokyo, Japan.
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Miki
authors.affiliation: Department of Molecular Genetics; Medical Research Institute; Tokyo Medical and Dental University (TMDU); Bunkyo-ku, Tokyo, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Nakanishi
mesh_terms: D002455:Cell Division / Q000502:physiology*; D045744:Cell Line, Tumor; D018385:Centrosome / Q000378:metabolism*; D006801:Humans; D025821:Polyubiquitin / Q000378:metabolism; D016196:S Phase / Q000502:physiology; D057149:Ubiquitinated Proteins / Q000378:metabolism*
chemical_list: D057149:Ubiquitinated Proteins; D025821:Polyubiquitin
keywords: M phase; cell cycle; centrosome; maturation; proteasome; proteome; ubiquitination
references.citation: Biochemistry. 2009 Sep 8;48(35):8483-90
references.pmid: 19642705
references.citation: Cell Mol Life Sci. 2013 Feb;70(4):661-87
references.pmid: 22864622
references.citation: Cancer Res. 2004 Aug 1;64(15):5106-12
references.pmid: 15289312
references.citation: Proc Natl Acad Sci U S A. 2010 Dec 7;107(49):21022-7
references.pmid: 21097701
references.citation: J Biol Chem. 2012 Jan 20;287(4):2317-27
references.pmid: 22069321
references.citation: J Cell Biol. 1999 May 3;145(3):481-90
references.pmid: 10225950
references.citation: Biochem Biophys Res Commun. 2006 Dec 22;351(3):658-63
references.pmid: 17083915
references.citation: J Cell Biol. 2012 Apr 2;197(1):19-26
references.pmid: 22451695
references.citation: J Biol Chem. 2010 Oct 8;285(41):31616-33
references.pmid: 20682791
references.citation: J Biol Chem. 2000 Jan 7;275(1):409-13
references.pmid: 10617632
references.citation: Nature. 2003 Dec 4;426(6966):570-4
references.pmid: 14654843
references.citation: DNA Repair (Amst). 2012 Feb 1;11(2):157-66
references.pmid: 22055568
references.citation: Oncogene. 2008 Aug 21;27(36):4900-8
references.pmid: 18427546
references.citation: Oncogene. 2008 Mar 13;27(12):1665-76
references.pmid: 17891179
references.citation: J Biol Chem. 2012 Mar 16;287(12):8803-15
references.pmid: 22270370
references.citation: J Biol Chem. 2011 Oct 21;286(42):36641-51
references.pmid: 21878651
references.citation: Commun Integr Biol. 2011 May;4(3):308-11
references.pmid: 21980565
references.citation: Int J Biochem Cell Biol. 2008;40(10):2315-32
references.pmid: 18487071
references.citation: Mol Cell Biol. 2007 Jan;27(1):352-67
references.pmid: 17060449
references.citation: Mol Biol Cell. 2008 Mar;19(3):1220-9
references.pmid: 18094058
references.citation: Cell. 2003 Dec 12;115(6):727-38
references.pmid: 14675537
references.citation: Proc Natl Acad Sci U S A. 2008 Oct 28;105(43):16743-8
references.pmid: 18948594
references.citation: J Biol Chem. 2011 Oct 21;286(42):36652-66
references.pmid: 21878652
references.citation: J Biol Chem. 2009 Aug 21;284(34):22970-7
references.pmid: 19509300
references.citation: Cell. 2009 Apr 3;137(1):133-45
references.pmid: 19345192
references.citation: Mol Cell. 2012 Sep 14;47(5):694-706
references.pmid: 22864114
references.citation: J Mol Biol. 2010 Jan 22;395(3):587-94
references.pmid: 19913553
references.citation: J Cell Biol. 2010 Apr 19;189(2):261-74
references.pmid: 20404110
references.citation: Cell Cycle. 2008 Jul 15;7(14):2225-33
references.pmid: 18635967
references.citation: Dev Cell. 2013 May 13;25(3):229-40
references.pmid: 23643362
references.citation: Dev Cell. 2003 Jul;5(1):113-25
references.pmid: 12852856
references.citation: Oncogene. 2011 May 12;30(19):2188-97
references.pmid: 21242972
references.citation: J Cell Biol. 2011 Dec 26;195(7):1093-101
references.pmid: 22184200
references.citation: J Cell Biol. 1998 Dec 28;143(7):1883-98
references.pmid: 9864362
references.citation: Proc Natl Acad Sci U S A. 2008 Dec 23;105(51):20197-202
references.pmid: 19091944
references.citation: Trends Biochem Sci. 2012 Dec;37(12):534-42
references.pmid: 23141205
references.citation: EMBO J. 2001 Jun 1;20(11):2878-84
references.pmid: 11387220
references.citation: Nat Cell Biol. 2003 Jun;5(6):505-11
references.pmid: 12776127