SUMOylated NKAP is essential for chromosome alignment by anchoring CENP-E to kinetochores.
Chromosome alignment is required for accurate chromosome segregation. Chromosome misalignment can result in genomic instability and tumorigenesis. Here, we show that NF-κB activating protein (NKAP) is critical for chromosome alignment through anchoring CENP-E to kinetochores. NKAP knockdown causes chromosome misalignment and prometaphase arrest in human cells. NKAP dynamically localizes to kinetochores, and is required for CENP-E kinetochore localization. NKAP is SUMOylated predominantly in mitosis and the SUMOylation is needed for NKAP to bind CENP-E. A SUMOylation-deficient mutant of NKAP cannot support the localization of CENP-E on kinetochores or proper chromosome alignment. Moreover, Bub3 recruits NKAP to stabilize the binding of CENP-E to BubR1 at kinetochores. Importantly, loss of NKAP expression causes aneuploidy in cultured cells, and is observed in human soft tissue sarcomas. These findings indicate that NKAP is a novel and key regulator of mitosis, and its dysregulation might contribute to tumorigenesis by causing chromosomal instability.
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Teng
T
Li
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Liang
L
Chen
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Juanxian
J
Cheng
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Jiang
J
Dai
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Yijiao
Y
Huang
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Jian
J
Zhang
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Zhaoshan
Z
Liu
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Ang
A
Li
Department of Gynecology, Affiliated Hospital of Academy of Military Medical Sciences, Beijing 100071, China.
Na
N
Li
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Hongxia
H
Wang
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Xiaomin
X
Yin
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Kun
K
He
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Ming
M
Yu
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Tao
T
Zhou
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Xuemin
X
Zhang
State Key Laboratory of Proteomics, Institute of Basic Medical Sciences, National Center of Biomedical Analysis, 27 Tai-Ping Road, Beijing 100850, China.
Qing
Q
Xia
D000782:Aneuploidy; D063646:Carcinogenesis; D018797:Cell Cycle Proteins / Q000378:metabolism; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism*; D002875:Chromosomes / Q000737:chemistry / Q000648:ultrastructure*; D056970:Co-Repressor Proteins / Q000378:metabolism*; D015972:Gene Expression Regulation, Neoplastic; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000737:chemistry*; D008938:Mitosis; D009154:Mutation; D009687:Nuclear Proteins / Q000378:metabolism*; D000075223:Poly-ADP-Ribose Binding Proteins / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D012097:Repressor Proteins; D012509:Sarcoma / Q000378:metabolism; D058207:Sumoylation
C489444:BUB3 protein, human; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D056970:Co-Repressor Proteins; C479077:NKAP protein, human; D009687:Nuclear Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D012097:Repressor Proteins; C068688:centromere protein E; C584277:BUB1 protein, human; D017346:Protein Serine-Threonine Kinases
Nat Rev Mol Cell Biol. 2007 May;8(5):379-93
17426725
Nature. 2010 Apr 1;464(7289):721-7
20360735
J Cell Biol. 1999 Sep 6;146(5):941-54
10477750
Nat Cell Biol. 2009 Jan;11(1):27-35
19060894
Mol Biol Cell. 2001 Sep;12(9):2776-89
11553716
J Cell Biol. 1998 Oct 5;143(1):49-63
9763420
Nucleic Acids Res. 2014 Mar;42(5):3177-93
24353314
Nat Cell Biol. 2011 Jul 10;13(8):924-33
21743465
Nat Cell Biol. 2013 Sep;15(9):1079-88
23955301
Nat Commun. 2014 Nov 13;5:5409
25391492
Biochem Biophys Res Commun. 2003 Oct 24;310(3):720-4
14550261
Curr Biol. 2010 Mar 23;20(6):R285-95
20334839
Nat Rev Mol Cell Biol. 2010 Dec;11(12):861-71
21102611
Nat Struct Mol Biol. 2014 Oct;21(10):927-36
25218447
J Cell Biol. 2003 Aug 18;162(4):551-63
12925705
J Cell Sci. 2004 Mar 15;117(Pt 8):1577-89
15020684
Cell Cycle. 2010 Mar 15;9(6):1207-13
20237434
Nat Rev Cancer. 2010 Feb;10(2):102-15
20094045
Nat Cell Biol. 2014 Dec;16(12 ):1249-56
25383660
Cell. 2008 Dec 12;135(6):1039-52
19070575
Chromosoma. 1997 Dec;106(7):446-55
9391217
Nat Rev Mol Cell Biol. 2013 Jan;14(1):25-37
23258294
Nat Rev Mol Cell Biol. 2010 Feb;11(2):91-102
20068571
Sci Signal. 2009 May 26;2(72):ra24
19471022
J Cell Biol. 2008 May 5;181(3):411-9
18443223
Mol Cell. 2008 Mar 28;29(6):729-41
18374647
Nat Cell Biol. 2015 Sep;17 (9):1124-33
26258632
J Biol Chem. 2007 Jul 20;282(29):21415-24
17535814
Nat Rev Mol Cell Biol. 2008 Jan;9(1):33-46
18097444
Nat Biotechnol. 2011 Jan;29(1):24-6
21221095
J Cell Biol. 2015 Apr 27;209(2):221-34
25918224
Dev Cell. 2014 Feb 10;28(3):268-81
24462186
Nat Rev Mol Cell Biol. 2014 Nov;15(11):736-47
25303117
Cell Cycle. 2010 Feb 15;9(4):715-9
20160491
Nat Cell Biol. 2000 Aug;2(8):484-91
10934468
Mol Cell Biol. 2005 May;25(10 ):4062-74
15870278
Cell. 2003 Jul 11;114(1):87-98
12859900
EMBO J. 1991 May;10(5):1245-54
2022189
Cancer Cell. 2007 Jan;11(1):25-36
17189716
Science. 2002 Sep 27;297(5590):2267-70
12351790
Immunity. 2009 May;30(5):696-707
19409814
Cold Spring Harb Perspect Biol. 2014 Jul 01;6(7):a015826
24984773
Genes Dev. 2004 Sep 15;18(18):2255-68
15371340
Science. 2006 Jan 20;311(5759):388-91
16424343
Sci Signal. 2010 Jan 12;3(104):ra3
20068231
Nat Rev Mol Cell Biol. 2009 Jul;10(7):478-87
19546858
Sci Signal. 2014 Apr 29;7(323):rs2
24782567
Cell. 2013 Feb 28;152(5):1160-72
23452859
Trends Cell Biol. 2005 Aug;15(8):442-51
16023855
Mol Biol Cell. 2008 Apr;19(4):1798-813
18199686
Dev Cell. 2002 Sep;3(3):351-65
12361599
J Cell Biol. 1997 Jun 30;137(7):1567-80
9199171