Temporal changes in Hec1 phosphorylation control kinetochore-microtubule attachment stability during mitosis.
Precise control of the attachment strength between kinetochores and spindle microtubules is essential to preserve genomic stability. Aurora B kinase has been implicated in regulating the stability of kinetochore-microtubule attachments but its relevant kinetochore targets in cells remain unclear. Here, we identify multiple serine residues within the N-terminus of the kinetochore protein Hec1 that are phosphorylated in an Aurora-B-kinase-dependent manner during mitosis. On all identified target sites, Hec1 phosphorylation at kinetochores is high in early mitosis and decreases significantly as chromosomes bi-orient. Furthermore, once dephosphorylated, Hec1 is not highly rephosphorylated in response to loss of kinetochore-microtubule attachment or tension. We find that a subpopulation of Aurora B kinase remains localized at the outer kinetochore even upon Hec1 dephosphorylation, suggesting that Hec1 phosphorylation by Aurora B might not be regulated wholly by spatial positioning of the kinase. Our results define a role for Hec1 phosphorylation in kinetochore-microtubule destabilization and error correction in early mitosis and for Hec1 dephosphorylation in maintaining stable attachments in late mitosis.
Department of Biochemistry and Molecular Biology, Colorado State University, Fort Collins, CO 80523, USA.
Keith F
KF
DeLuca
Susanne M A
SM
Lens
Jennifer G
JG
DeLuca
D064107:Aurora Kinase B; D064127:Aurora Kinases; D003598:Cytoskeletal Proteins; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000737:chemistry / Q000378:metabolism*; D008870:Microtubules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008938:Mitosis; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D055550:Protein Stability
D003598:Cytoskeletal Proteins; C120529:NDC80 protein, human; D009687:Nuclear Proteins; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases
Cell. 2002 Feb 8;108(3):317-29
11853667
Science. 1999 Oct 29;286(5441):971-4
10542155
Science. 2002 Sep 27;297(5590):2267-70
12351790
Cell. 2002 Oct 18;111(2):163-72
12408861
Science. 2009 Mar 6;323(5919):1350-3
19150808
Dev Cell. 2008 Feb;14(2):252-62
18267093
Dev Cell. 2004 Feb;6(2):253-68
14960279
J Cell Biol. 2001 Jan 22;152(2):349-60
11266451
Mol Cell. 2005 Apr 29;18(3):379-91
15866179
Mol Cell Proteomics. 2004 Jan;3(1):93-104
14602875
Cell. 2008 May 2;133(3):427-39
18455984
Curr Biol. 2008 Nov 25;18(22):1785-91
19026542
J Cell Biol. 2003 Apr 28;161(2):281-94
12707311
J Cell Biol. 2000 Sep 4;150(5):975-88
10973989
Nat Cell Biol. 2005 Jan;7(1):93-8
15592459
Mol Cell. 2010 May 14;38(3):383-92
20471944
Genes Dev. 2001 Dec 1;15(23):3118-29
11731476
Curr Biol. 2004 Jan 20;14(2):131-7
14738735
Nat Cell Biol. 2009 Jan;11(1):27-35
19060894
J Cell Biol. 2002 Nov 25;159(4):549-55
12438418
Sci Signal. 2010 Jan 12;3(104):ra3
20068231
Curr Biol. 2004 Dec 14;14(23):2149-55
15589159
Curr Biol. 2008 Nov 25;18(22):1778-84
19026543
Proc Natl Acad Sci U S A. 2006 Apr 4;103(14):5391-6
16565220
Curr Biol. 2006 Sep 5;16(17):1711-8
16950108
Curr Opin Cell Biol. 2006 Dec;18(6):623-31
17030123
Cell. 2006 Dec 1;127(5):983-97
17129783
J Cell Biol. 2003 Apr 28;161(2):267-80
12719470
Mol Cell Biol. 1997 Oct;17(10):6049-56
9315664
J Proteome Res. 2009 Oct;8(10):4553-63
19691289
Genetics. 2009 Dec;183(4):1591-5
19822728
J Cell Biol. 2010 Mar 22;188(6):809-20
20231380
Curr Biol. 2006 Sep 5;16(17):1705-10
16950107
Cell Div. 2008 May 28;3:10
18507820
Science. 1996 Oct 11;274(5285):242-6
8824188
J Biol Chem. 2004 Mar 26;279(13):12997-3003
14722118
Cell Stem Cell. 2009 Aug 7;5(2):204-13
19664994
Cell. 2006 Dec 1;127(5):969-82
17129782
Nat Struct Mol Biol. 2007 Jan;14(1):54-9
17195848
J Cell Biol. 1995 Nov;131(3):721-34
7593192
Science. 1996 Oct 11;274(5285):246-8
8824189
Biochem Soc Trans. 1978;6(1):17-20
205460
Nat Cell Biol. 2006 Jan;8(1):78-83
16327780
Biochim Biophys Acta. 2008 Sep;1786(1):32-40
18549824