Centrosome biogenesis continues in the absence of microtubules during prolonged S-phase arrest.
When CHO cells are arrested in S-phase, they undergo repeated rounds of centrosome duplication without cell-cycle progression. While the increase is slow and asynchronous, the number of centrosomes in these cells does rise with time. To investigate mechanisms controlling this duplication, we have arrested CHO cells in S-phase for up to 72 h, and coordinately inhibited new centriole formation by treatment with the microtubule poison colcemid. We find that in such cells, the pre-existing centrosomes remain, and a variable number of foci--containing alpha/gamma-tubulin and centrin 2--assemble at the nuclear periphery. When the colcemid is washed out, the nuclear-associated foci disappear, and cells assemble new centriole-containing centrosomes, which accumulate the centriole scaffold protein SAS-6, nucleate microtubule asters, and form functional mitotic spindle poles. The number of centrosomes that assemble following colcemid washout increases with duration of S-phase arrest, even though the number of nuclear-associated foci or pre-existing centrosomes does not increase. This suggests that during S-phase, a cryptic generative event occurs repeatedly, even in the absence of new triplet microtubule assembly. When triplet microtubule assembly is restored, these cryptic generative events become realized, and multiple centriole-containing centrosomes assemble.
Collins
Elizabeth S
ES
Cellular Dynamics Section, The Hormel Institute, University of Minnesota, Austin, Minnesota, USA.
Hornick
Jessica E
JE
Durcan
Thomas M
TM
Collins
Nicholas S
NS
Archer
William
W
Karanjeet
Kul B
KB
Vaughan
Kevin T
KT
Hinchcliffe
Edward H
EH
D000818:Animals; D016466:CHO Cells; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D018385:Centrosome / Q000187:drug effects / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D003703:Demecolcine / Q000494:pharmacology; D015200:Fluoroimmunoassay; D005786:Gene Expression Regulation / Q000502:physiology; D049452:Green Fluorescent Proteins; D006918:Hydroxyurea / Q000494:pharmacology; D008870:Microtubules / Q000502:physiology*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D050257:Tubulin Modulators / Q000494:pharmacology
D018797:Cell Cycle Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D050257:Tubulin Modulators; D049452:Green Fluorescent Proteins; D006918:Hydroxyurea; D003703:Demecolcine
J Cell Biol. 1998 Mar 23;140(6):1417-26
9508774
Curr Biol. 2009 Jan 13;19(1):43-9
19084407
J Cell Biol. 2000 Apr 17;149(2):317-30
10769025
Curr Biol. 2007 Dec 18;17(24):2169-74
18082404
Mol Cell Biol. 2009 Apr;29(7):1760-73
19139275
J Cell Biol. 1975 Apr;65(1):65-74
805149
Curr Biol. 1999 Apr 22;9(8):429-32
10226033
Methods Cell Biol. 2007;81:315-33
17519173
Genes Dev. 2001 May 15;15(10):1167-81
11358861
Curr Biol. 2007 May 15;17(10):834-43
17475495
J Cell Biol. 1967 Aug;34(2):525-33
6068183
J Cell Biol. 2002 Sep 30;158(7):1171-81
12356862
J Cell Sci. 1982 Feb;53:155-71
7200989
J Cell Sci. 2004 Apr 1;117(Pt 9):1619-30
15075224
Microsc Res Tech. 2000 Jun 1;49(5):428-34
10842369
J Exp Zool. 1987 Oct;244(1):89-100
3694143
Cell Motil Cytoskeleton. 1996;33(1):1-5
8824729
Biol Cell. 1991;72(1-2):31-8
1756310
J Cell Biol. 1998 Dec 14;143(6):1575-89
9852152
J Cell Biol. 1977 Jun;73(3):601-15
559676
J Cell Sci. 2007 Jul 15;120(Pt 14):2444-53
17606999
Nat Cell Biol. 2007 Jul;9(7):736-8
17603508
Genes Dev. 1999 Sep 1;13(17):2242-57
10485847
J Biophys Biochem Cytol. 1961 Oct;11:244-7
13886557
Curr Biol. 2001 Mar 6;11(5):308-17
11267867
J Cell Biol. 2001 Apr 2;153(1):237-42
11285289
J Cell Biol. 1981 Dec;91(3 Pt 1):814-21
7328123
Cell Motil Cytoskeleton. 2008 Aug;65(8):595-613
18481305
J Cell Physiol. 2008 Apr;215(1):182-91
17960592
Cell Motil Cytoskeleton. 2000 Sep;47(1):13-24
11002307
Nat Cell Biol. 2003 Jun;5(6):539-44
12766773
J Cell Biol. 1990 May;110(5):1599-605
2335566
J Cell Biol. 2005 Feb 28;168(5):713-22
15738265
Science. 1999 Feb 5;283(5403):851-4
9933170
Nat Rev Mol Cell Biol. 2007 Jun;8(6):451-63
17505520
J Cell Biol. 2009 Jan 26;184(2):225-39
19171756
J Cell Biol. 2007 Jan 15;176(2):173-82
17227892
Curr Biol. 2009 Jun 23;19(12):1012-8
19481460
Science. 1992 Apr 10;256(5054):219-21
1566068
Curr Biol. 2005 Dec 20;15(24):2199-207
16326102
Methods Cell Biol. 1999;61:297-315
9891321
J Cell Biol. 1995 Jul;130(1):105-15
7790366
J Cell Biol. 1990 Jun;110(6):2033-42
2190990
Curr Biol. 2003 Jan 21;13(2):R55-6
12546804
Nature. 2006 Nov 30;444(7119):619-23
17136092
Oncogene. 2007 Sep 20;26(43):6280-8
17438528
J Cell Biol. 2008 May 19;181(4):595-603
18474621
Cell. 2008 Jun 13;133(6):1032-42
18555779
Proc Natl Acad Sci U S A. 1968 Oct;61(2):461-8
4176480
Int J Cancer. 2006 Dec 15;119(12):2717-23
17016823
J Cell Biol. 1990 Jun;110(6):2025-32
2351692
J Gen Virol. 1974 Aug;24(2):395-9
4853583
Dev Cell. 2007 Aug;13(2):203-13
17681132
Science. 2007 May 18;316(5827):1046-50
17463247
Cell Motil Cytoskeleton. 1999;42(1):60-72
9915585
Curr Opin Cell Biol. 2004 Feb;16(1):49-54
15037304
Nat Cell Biol. 2005 Nov;7(11):1140-6
16244668
Science. 2001 Feb 23;291(5508):1547-50
11222860
J Cell Biol. 1999 Aug 9;146(3):585-96
10444067
Curr Biol. 2002 Aug 6;12(15):1287-92
12176356
Nature. 2006 Aug 24;442(7105):947-51
16862117
Curr Biol. 2007 Dec 4;17(23):2081-6
18060787