title: CENP-32 is required to maintain centrosomal dominance in bipolar spindle assembly.
abstract: Centrosomes nucleate spindle formation, direct spindle pole positioning, and are important for proper chromosome segregation during mitosis in most animal cells. We previously reported that centromere protein 32 (CENP-32) is required for centrosome association with spindle poles during metaphase. In this study, we show that CENP-32 depletion seems to release centrosomes from bipolar spindles whose assembly they had previously initiated. Remarkably, the resulting anastral spindles function normally, aligning the chromosomes to a metaphase plate and entering anaphase without detectable interference from the free centrosomes, which appear to behave as free asters in these cells. The free asters, which contain reduced but significant levels of CDK5RAP2, show weak interactions with spindle microtubules but do not seem to make productive attachments to kinetochores. Thus CENP-32 appears to be required for centrosomes to integrate into a fully functional spindle that not only nucleates astral microtubules, but also is able to nucleate and bind to kinetochore and central spindle microtubules. Additional data suggest that NuMA tethers microtubules at the anastral spindle poles and that augmin is required for centrosome detachment after CENP-32 depletion, possibly due to an imbalance of forces within the spindle.
authors: Wellcome Trust Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh EH9 3JR, United Kingdom Medical School, Kochi University, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan shinya.ohta@kochi-u.ac.jp bill.earnshaw@ed.ac.uk.
authors: Shinya
authors: S
authors: Ohta
authors: Wellcome Trust Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh EH9 3JR, United Kingdom.
authors: Laura
authors: L
authors: Wood
authors: Medical School, Kochi University, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan.
authors: Iyo
authors: I
authors: Toramoto
authors: Medical School, Kochi University, Kohasu, Oko-cho, Nankoku, Kochi 783-8505, Japan.
authors: Ken-Ichi
authors: K
authors: Yagyu
authors: Department of Molecular Genetics, National Institute of Genetics and the Graduate University for Advanced Studies, Mishima, Shizuoka 411-8540, Japan.
authors: Tatsuo
authors: T
authors: Fukagawa
authors: Wellcome Trust Centre for Cell Biology, School of Biological Sciences, University of Edinburgh, Edinburgh EH9 3JR, United Kingdom shinya.ohta@kochi-u.ac.jp bill.earnshaw@ed.ac.uk.
authors: William C
authors: WC
authors: Earnshaw
mesh_terms: D034961:Antigens, Nuclear / Q000378:metabolism; D018797:Cell Cycle Proteins; D018385:Centrosome / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D006801:Humans; D018386:Kinetochores / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D034521:Nuclear Matrix-Associated Proteins / Q000378:metabolism; D008941:Spindle Apparatus / Q000378:metabolism*
chemical_list: D034961:Antigens, Nuclear; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; C050045:NUMA1 protein, human; D034521:Nuclear Matrix-Associated Proteins; C000606699:SPOUT1 protein, human
references: Cold Spring Harb Symp Quant Biol. 1974;38:123-31
references: 4598635
references: Nat Cell Biol. 2005 Jan;7(1):42-7
references: 15592460
references: J Cell Sci. 2009 Jul 15;122(Pt 14):2543-53
references: 19549687
references: Nature. 1992 Oct 8;359(6395):540-3
references: 1406972
references: J Cell Biol. 2008 May 5;181(3):421-9
references: 18443220
references: Mol Biol Cell. 2013 Oct;24(20):3238-50
references: 23966467
references: J Cell Biol. 2001 Dec 24;155(7):1109-16
references: 11748251
references: J Biol Chem. 1999 Jun 11;274(24):17267-74
references: 10358086
references: BMC Res Notes. 2009 Apr 28;2:64
references: 19400937
references: Cytoskeleton (Hoboken). 2012 Mar;69(3):155-65
references: 22278908
references: Neurogastroenterol Motil. 2009 Aug;21(8):813-e52
references: 19368661
references: Cell. 2010 Sep 3;142(5):810-21
references: 20813266
references: Dev Cell. 2003 Jul;5(1):113-25
references: 12852856
references: Plant J. 2013 Jul;75(2):245-57
references: 23521421
references: Proc Natl Acad Sci U S A. 2009 Apr 28;106(17):6998-7003
references: 19369198
references: Nat Rev Mol Cell Biol. 2007 Feb;8(2):161-7
references: 17245416
references: Nat Cell Biol. 2012 Mar;14(3):311-7
references: 22327364
references: J Cell Biol. 1995 Nov;131(3):693-708
references: 7593190
references: Nat Cell Biol. 2006 Oct;8(10):1095-101
references: 16980960
references: EMBO J. 2009 Jul 22;28(14):2066-76
references: 19536135
references: Curr Biol. 2000 Nov 2;10(21):1367-70
references: 11084336
references: Mol Biol Cell. 2000 Jan;11(1):241-53
references: 10637305
references: J Cell Biol. 2000 May 15;149(4):851-62
references: 10811826
references: Curr Biol. 1998 Jul 30-Aug 13;8(16):903-13
references: 9707401
references: Plant Cell. 2011 Jul;23(7):2606-18
references: 21750235
references: J Cell Biol. 1998 Nov 2;143(3):687-94
references: 9813090
references: J Biol Chem. 1997 May 23;272(21):13766-71
references: 9153231
references: Science. 2001 Jan 26;291(5504):653-6
references: 11229403
references: Plant Cell. 2012 Apr;24(4):1494-509
references: 22505726
references: Cell. 1992 Aug 7;70(3):451-8
references: 1643659
references: Science. 1999 Oct 29;286(5441):971-4
references: 10542155
references: Mol Biol Cell. 2008 Jan;19(1):115-25
references: 17959831
references: Mol Cell Biol. 2000 Oct;20(20):7813-25
references: 11003675
references: Dev Biol. 2012 Aug 15;368(2):242-54
references: 22613359
references: J Cell Sci. 1999 Jul;112 ( Pt 14):2453-61
references: 10381400
references: J Cell Biol. 2013 Jul 8;202(1):25-33
references: 23816620
references: Proc Natl Acad Sci U S A. 2013 Dec 3;110(49):19808-13
references: 24255106
references: Cell. 1996 Nov 1;87(3):447-58
references: 8898198
references: Mol Biol Cell. 2013 Aug;24(16):2506-14
references: 23783028
references: Science. 1999 May 21;284(5418):1359-62
references: 10334991
references: J Cell Biol. 2005 Jan 3;168(1):141-53
references: 15631994
references: Curr Opin Plant Biol. 2013 Dec;16(6):698-703
references: 24075308
references: PLoS One. 2013;8(7):e68523
references: 23874654
references: Cell. 1986 May 9;45(3):329-42
references: 3516413
references: J Cell Biol. 1985 Sep;101(3):766-77
references: 4030894
references: Nat Cell Biol. 2003 Feb;5(2):143-8
references: 12524548
references: Nat Cell Biol. 2011 Sep;13(9):1040-50
references: 21822276
references: Science. 2002 Mar 29;295(5564):2452-6
references: 11923538
references: J Photochem Photobiol B. 1995 Aug;29(2-3):163-70
references: 7472810
references: J Cell Biol. 2010 Apr 5;189(1):23-39
references: 20368616
references: Dev Cell. 2007 Apr;12(4):531-41
references: 17419992
references: Mol Biol Cell. 1997 Jun;8(6):1025-33
references: 9201713
references: J Cell Biol. 2012 Sep 17;198(6):1039-54
references: 22965910
references: Mol Biol Cell. 2005 Jul;16(7):3176-86
references: 15888542
references: Mol Biol Cell. 2000 May;11(5):1765-74
references: 10793150
references: Cell. 2012 Feb 3;148(3):502-14
references: 22304918
references: Mol Cell. 2012 Sep 14;47(5):694-706
references: 22864114
references: Annu Rev Cell Dev Biol. 2005;21:411-34
references: 16212501
references: Mol Biol Cell. 2002 Sep;13(9):3235-45
references: 12221128
references: J Biol Chem. 2000 Nov 3;275(44):34592-6
references: 10945988
references: J Cell Biol. 1998 Apr 6;141(1):163-74
references: 9531556
references: J Cell Biol. 1984 Jul;99(1 Pt 2):47s-54s
references: 6235234
references: Dev Cell. 2010 Jun 15;18(6):913-26
references: 20627074
references: Nature. 1996 Aug 1;382(6590):420-5
references: 8684481
references: Dev Cell. 2007 Apr;12(4):515-29
references: 17419991
references: Nat Cell Biol. 2001 Mar;3(3):221-7
references: 11231570
references: Cell. 2013 Jul 18;154(2):391-402
references: 23870127
references: Curr Biol. 2000 Jan 27;10(2):59-67
references: 10662665
references: J Cell Sci. 2007 May 15;120(Pt 10):1717-22
references: 17502482
references: J Cell Biol. 2010 Oct 18;191(2):259-67
references: 20937700