Proteomic and functional analysis of the mitotic Drosophila centrosome.
Regulation of centrosome structure, duplication and segregation is integrated into cellular pathways that control cell cycle progression and growth. As part of these pathways, numerous proteins with well-established non-centrosomal localization and function associate with the centrosome to fulfill regulatory functions. In turn, classical centrosomal components take up functional and structural roles as part of other cellular organelles and compartments. Thus, although a comprehensive inventory of centrosome components is missing, emerging evidence indicates that its molecular composition reflects the complexity of its functions. We analysed the Drosophila embryonic centrosomal proteome using immunoisolation in combination with mass spectrometry. The 251 identified components were functionally characterized by RNA interference. Among those, a core group of 11 proteins was critical for centrosome structure maintenance. Depletion of any of these proteins in Drosophila SL2 cells resulted in centrosome disintegration, revealing a molecular dependency of centrosome structure on components of the protein translation machinery, actin- and RNA-binding proteins. In total, we assigned novel centrosome-related functions to 24 proteins and confirmed 13 of these in human cells.
Müller
Hannah
H
Department of Vertebrate Genomics, Max-Planck Institute for Molecular Genetics, Berlin, Germany.
Schmidt
David
D
Steinbrink
Sandra
S
Mirgorodskaya
Ekaterina
E
Lehmann
Verena
V
Habermann
Karin
K
Dreher
Felix
F
Gustavsson
Niklas
N
Kessler
Thomas
T
Lehrach
Hans
H
Herwig
Ralf
R
Gobom
Johan
J
Ploubidou
Aspasia
A
Boutros
Michael
M
Lange
Bodo M H
BM
D000818:Animals; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D018385:Centrosome / Q000737:chemistry* / Q000502:physiology; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000378:metabolism*; D004330:Drosophila / Q000737:chemistry* / Q000502:physiology; D004625:Embryo, Nonmammalian / Q000378:metabolism / Q000502:physiology; D013058:Mass Spectrometry; D008938:Mitosis / Q000502:physiology*; D040901:Proteomics / Q000379:methods; D034622:RNA Interference
D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone
Trends Cell Biol. 1993 Apr;3(4):118-28
14731766
Genes Dev. 1999 Jun 1;13(11):1422-37
10364159
Oncogene. 2008 Nov 24;27(55):6994-7002
19029940
Proteomics. 2004 Sep;4(9):2707-21
15352245
J Cell Biol. 2009 Nov 2;187(3):355-63
19948479
Genes Dev. 2001 May 15;15(10):1167-81
11358861
Curr Biol. 2008 Feb 26;18(4):303-9
18291647
Nat Rev Mol Cell Biol. 2001 Sep;2(9):688-98
11533726
J Cell Biol. 1995 Sep;130(5):1149-59
7657699
PLoS Biol. 2008 Apr 22;6(4):e98
18433294
J Cell Biol. 1998 May 18;141(4):967-77
9585415
J Cell Sci. 2004 Apr 15;117(Pt 10):1911-22
15090595
Dev Biol. 2010 Apr 15;340(2):408-18
20122915
Development. 2004 Feb;131(4):863-71
14736750
Nature. 2004 Feb 19;427(6976):753-7
14973490
J Cell Biol. 2008 Feb 25;180(4):681-9
18283114
Proc Natl Acad Sci U S A. 1998 Mar 17;95(6):2950-5
9501196
Cell. 2006 May 5;125(3):523-33
16678096
Curr Opin Cell Biol. 2002 Feb;14(1):25-34
11792541
J Cell Sci. 1996 Dec;109 ( Pt 13):3001-12
9004035
Curr Biol. 2007 Sep 4;17(17):1465-72
17689959
J Cell Biol. 1999 Feb 22;144(4):721-33
10037793
J Cell Biol. 2000 Jul 24;150(2):F37-44
10908584
Mol Cell Biol. 2008 Mar;28(5):1755-69
18172010
Mol Cell Biol. 1998 Jun;18(6):3540-51
9584194
Gene. 2004 Feb 4;326:107-16
14729268
Neuron. 2003 Sep 25;40(1):25-40
14527431
J Cell Biol. 1985 Jan;100(1):35-46
3880758
Nucleic Acids Res. 2009 Jan;37(Database issue):D175-80
18971254
J Cell Sci. 1997 Feb;110 ( Pt 4):451-64
9067597
J Cell Biol. 2010 Feb 8;188(3):313-23
20123993
EMBO J. 2000 Mar 15;19(6):1252-62
10716925
Mol Biol Cell. 2009 May;20(10):2605-14
19321663
Nat Rev Genet. 2005 Mar;6(3):194-205
15738963
J Cell Biol. 2003 Dec 22;163(6):1197-204
14691132
Curr Biol. 2007 Oct 23;17(20):1735-45
17935995
J Biol Chem. 2004 Apr 2;279(14):13721-8
14739278
Cell. 2005 Apr 22;121(2):223-34
15851029
Mol Cell Biol. 1996 Dec;16(12):6870-8
8943342
J Cell Biol. 2004 Jun 7;165(5):673-83
15184400
J Biol Chem. 2009 Apr 3;284(14):9418-25
19201755
J Cell Biol. 2001 Apr 2;153(1):237-42
11285289
Nat Rev Cancer. 2002 Nov;2(11):815-25
12415252
Nat Cell Biol. 2001 Jul;3(7):643-9
11433296
Curr Opin Cell Biol. 2002 Feb;14(1):35-43
11792542
J Gen Virol. 2005 Mar;86(Pt 3):589-594
15722518
Biochem Biophys Res Commun. 2002 Aug 16;296(2):288-92
12163015
Curr Protoc Cell Biol. 2006 Jan;Chapter 3:Unit 3.17
18228485
Nat Cell Biol. 2001 Jan;3(1):68-75
11146628
Curr Biol. 2009 Jun 9;19(11):937-42
19427213
Trends Cell Biol. 2009 Jul;19(7):325-33
19576775
Genes Dev. 2008 Aug 15;22(16):2189-203
18662975
Science. 2006 Oct 27;314(5799):654-7
17068266
J Cell Sci. 2007 Dec 15;120(Pt 24):4321-31
18042621
Oncogene. 2002 Sep 9;21(40):6209-21
12214251
EMBO J. 2010 Feb 17;29(4):843-54
20111006
Genes Dev. 2000 Dec 15;14(24):3126-39
11124805
Curr Biol. 2007 Oct 23;17(20):1759-64
17919907
Nature. 2008 Dec 18;456(7224):971-5
19011615
Nat Rev Mol Cell Biol. 2007 Jun;8(6):451-63
17505520
Cell. 1996 May 17;85(4):585-96
8653793
J Cell Sci. 1994 Apr;107 ( Pt 4):859-67
8056842
Development. 1999 Jul;126(13):2829-39
10357928
J Cell Biol. 2006 Feb 13;172(4):517-28
16476773
J Cell Biol. 1989 Dec;109(6 Pt 1):2869-78
2592409
Nature. 2003 Dec 4;426(6966):570-4
14654843
Nat Cell Biol. 2010 Feb;12(2):164-9
20081840
Genes Dev. 1998 May 15;12(10):1483-94
9585508
J Cell Sci. 2005 Nov 15;118(Pt 22):5381-92
16280552
J Cell Biol. 2000 Jul 24;150(2):F51-6
10908586
Proteomics. 2005 Feb;5(2):399-408
15648048
J Cell Biol. 1995 Dec;131(5):1261-73
8522588
PLoS Genet. 2008 Jul 18;4(7):e1000126
18797514
Nat Cell Biol. 2007 Dec;9(12):1401-12
17994010
Mol Biol Cell. 1995 Nov;6(11):1563-74
8589456
Genes Dev. 2008 Jan 1;22(1):91-105
18086858
Science. 1966 Feb 25;151(3713):992-5
5907291
Cell. 2006 Jun 30;125(7):1375-86
16814722
Methods Cell Biol. 1999;61:35-56
9891308
Science. 2004 Feb 6;303(5659):832-5
14764878
Genetics. 2009 May;182(1):133-44
19293139
EMBO Rep. 2009 Jun;10(6):606-13
19390532
Nat Cell Biol. 2005 Oct;7(10):937-46
16184168
J Cell Biol. 2007 Aug 27;178(5):725-32
17709428
Mol Biol Cell. 2006 Jan;17(1):272-82
16236791
EMBO J. 2000 Jan 17;19(2):241-52
10637228
Science. 2007 Apr 20;316(5823):417-21
17412918
Curr Top Dev Biol. 2000;49:343-63
11005027
Genetics. 2008 Dec;180(4):2081-94
18854586
Science. 2007 May 18;316(5827):1046-50
17463247
PLoS Biol. 2008 Sep 16;6(9):e224
18798690
Development. 2002 Dec;129(24):5553-66
12421697
EMBO J. 2000 Aug 1;19(15):3932-44
10921875
Nat Cell Biol. 2000 Feb;2(2):90-5
10655588
Curr Top Dev Biol. 2000;49:449-70
11005031
Curr Biol. 2005 Jun 21;15(12):1090-8
15964273
Curr Biol. 2002 Aug 6;12(15):1287-92
12176356
Mol Biol Cell. 2004 Jan;15(1):37-45
14565985
J Cell Sci. 1997 Jul;110 ( Pt 13):1431-40
9224761
J Virol. 2006 Jun;80(12):5807-21
16731920
Cell. 2004 May 14;117(4):541-52
15137946