A genome-wide RNAi screen to dissect centriole duplication and centrosome maturation in Drosophila.
Centrosomes comprise a pair of centrioles surrounded by an amorphous pericentriolar material (PCM). Here, we have performed a microscopy-based genome-wide RNA interference (RNAi) screen in Drosophila cells to identify proteins required for centriole duplication and mitotic PCM recruitment. We analysed 92% of the Drosophila genome (13,059 genes) and identified 32 genes involved in centrosome function. An extensive series of secondary screens classified these genes into four categories: (1) nine are required for centriole duplication, (2) 11 are required for centrosome maturation, (3) nine are required for both functions, and (4) three genes regulate centrosome separation. These 32 hits include several new centrosomal components, some of which have human homologs. In addition, we find that the individual depletion of only two proteins, Polo and Centrosomin (Cnn) can completely block centrosome maturation. Cnn is phosphorylated during mitosis in a Polo-dependent manner, suggesting that the Polo-dependent phosphorylation of Cnn initiates centrosome maturation in flies.
Dobbelaere
Jeroen
J
The Gurdon Institute, Cambridge, United Kingdom.
Josué
Filipe
F
Suijkerbuijk
Saskia
S
Baum
Buzz
B
Tapon
Nicolas
N
Raff
Jordan
J
D000595:Amino Acid Sequence; D000818:Animals; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle / Q000502:physiology; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002502:Centrioles / Q000378:metabolism*; D018385:Centrosome / Q000378:metabolism* / Q000648:ultrastructure; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D004331:Drosophila melanogaster / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D016678:Genome; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D008941:Spindle Apparatus / Q000235:genetics / Q000378:metabolism
D029742:Caenorhabditis elegans Proteins; D018797:Cell Cycle Proteins; D029721:Drosophila Proteins; D018398:Homeodomain Proteins; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; C098130:cnn protein, Drosophila; C072004:polo protein, Drosophila; D017346:Protein Serine-Threonine Kinases
Curr Biol. 2002 Apr 16;12(8):640-7
11967150
J Biol Chem. 1999 Jan 22;274(4):2408-15
9891010
Dev Cell. 2007 Aug;13(2):190-202
17681131
Curr Biol. 2005 Dec 20;15(24):2199-207
16326102
Cell. 2006 Jun 30;125(7):1375-86
16814722
Mol Cell. 2005 Oct 28;20(2):237-49
16246726
J Cell Biol. 1996 Dec;135(6 Pt 2):1701-13
8991084
Curr Biol. 2007 May 15;17(10):834-43
17475495
Curr Biol. 2008 Feb 26;18(4):303-9
18291647
FEBS Lett. 1999 Jul 30;456(1):7-12
10452519
J Cell Biol. 1995 Jun;129(6):1617-28
7790358
Curr Opin Cell Biol. 2002 Feb;14(1):25-34
11792541
J Cell Biol. 1992 Jan;116(2):377-83
1309812
Curr Biol. 2007 Sep 4;17(17):1465-72
17689959
Nature. 2004 Sep 2;431(7004):92-6
15343338
Curr Biol. 2007 Oct 23;17(20):1735-45
17935995
FEBS Lett. 2001 Nov 9;508(1):136-42
11707284
J Cell Biol. 2001 Dec 24;155(7):1109-16
11748251
Curr Biol. 2004 May 25;14(10):863-73
15186742
J Cell Biol. 2004 Jun 7;165(5):673-83
15184400
Dev Cell. 2003 Mar;4(3):431-9
12636923
Mol Cell Biol. 2007 Jan;27(1):352-67
17060449
EMBO J. 1998 Jan 15;17(2):470-81
9430639
Curr Biol. 2008 Jan 22;18(2):136-41
18207742
Nat Cell Biol. 2001 Jan;3(1):68-75
11146628
Curr Opin Cell Biol. 1995 Feb;7(1):39-45
7755988
Cell. 1995 Apr 7;81(1):95-105
7720077
J Cell Biol. 2006 Feb 13;172(4):505-15
16461362
Curr Biol. 1999 Aug 26;9(16):889-98
10469591
Genes Dev. 2008 Mar 1;22(5):601-14
18316477
Cell. 2003 Feb 21;112(4):575-87
12600319
Proc Natl Acad Sci U S A. 2005 Aug 9;102(32):11325-30
16061793
Curr Biol. 2007 Oct 23;17(20):1759-64
17919907
Nat Rev Mol Cell Biol. 2007 Jun;8(6):451-63
17505520
Development. 1999 Jul;126(13):2829-39
10357928
Mol Biol Cell. 2008 Jul;19(7):3163-78
18463166
Annu Rev Biochem. 1994;63:639-74
7979251
Cell. 2001 May 18;105(4):547-58
11371350
J Cell Biol. 2006 Feb 13;172(4):517-28
16476773
Nature. 2003 Dec 4;426(6966):570-4
14654843
Dev Cell. 2004 Dec;7(6):815-29
15572125
Cancer Res. 2007 Nov 1;67(21):10103-5
17974949
Trends Cell Biol. 2007 May;17(5):215-21
17383880
Nat Cell Biol. 2006 Feb;8(2):137-47
16378099
Curr Opin Genet Dev. 2005 Jun;15(3):308-14
15917207
Nat Cell Biol. 2005 Feb;7(2):115-25
15665853
J Cell Biol. 2005 Oct 24;171(2):229-40
16247025
J Cell Biol. 2002 Jan 7;156(1):87-99
11781336
Mol Biol Cell. 2003 Mar;14(3):1017-26
12631720
Genome Biol. 2006;7(7):R66
16869968
Hum Mol Genet. 2005 Aug 1;14(15):2155-65
15972725
Cell. 2007 Aug 24;130(4):678-90
17719545
Trends Cell Biol. 2002 May;12(5):222-5
12062169
J Cell Biol. 2007 Aug 27;178(5):725-32
17709428
Cancer Cell. 2007 Jan;11(1):9-23
17189715
Cell. 2008 Jun 13;133(6):1032-42
18555779
Science. 2007 Apr 20;316(5823):417-21
17412918
Trends Cell Biol. 2002 Dec;12(12):551-5
12495842
Dev Cell. 2002 Sep;3(3):339-50
12361598
J Cell Biol. 2008 Feb 25;180(4):771-85
18299348
Nat Cell Biol. 2008 Mar;10(3):322-8
18297061
Curr Biol. 2007 Nov 20;17(22):1960-6
17980596
Development. 2002 Jan;129(2):347-59
11807028
Am J Hum Genet. 2005 May;76(5):717-28
15806441
J Cell Sci. 1988 Jan;89 ( Pt 1):25-38
3417791
Dev Cell. 2004 Apr;6(4):511-23
15068791
EMBO J. 1995 Jan 3;14(1):28-36
7828594
Nat Genet. 2005 Apr;37(4):353-5
15793586
Nature. 2006 Aug 24;442(7105):947-51
16862117
Annu Rev Cell Dev Biol. 2005;21:411-34
16212501
Nat Cell Biol. 2004 Jul;6(7):656-64
15232593
Genome Biol. 2006;7(10):R100
17076895