The constitutive centromere component CENP-50 is required for recovery from spindle damage.
We identified CENP-50 as a novel kinetochore component. We found that CENP-50 is a constitutive component of the centromere that colocalizes with CENP-A and CENP-H throughout the cell cycle in vertebrate cells. To determine the precise role of CENP-50, we examined its role in centromere function by generating a loss-of-function mutant in the chicken DT40 cell line. The CENP-50 knockout was not lethal; however, the growth rate of cells with this mutation was slower than that of wild-type cells. We observed that the time for CENP-50-deficient cells to complete mitosis was longer than that for wild-type cells. Centromeric localization of CENP-50 was abolished in both CENP-H- and CENP-I-deficient cells. Coimmunoprecipitation experiments revealed that CENP-50 interacted with the CENP-H/CENP-I complex in chicken DT40 cells. We also observed severe mitotic defects in CENP-50-deficient cells with apparent premature sister chromatid separation when the mitotic checkpoint was activated, indicating that CENP-50 is required for recovery from spindle damage.
Minoshima
Yukinori
Y
National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan.
Hori
Tetsuya
T
Okada
Masahiro
M
Kimura
Hiroshi
H
Haraguchi
Tokuko
T
Hiraoka
Yasushi
Y
Bao
Ying-Chun
YC
Kawashima
Toshiyuki
T
Kitamura
Toshio
T
Fukagawa
Tatsuo
T
D000595:Amino Acid Sequence; D000818:Animals; D018797:Cell Cycle Proteins; D002460:Cell Line; D002503:Centromere / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism*; D002645:Chickens; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020090:Chromosome Segregation / Q000187:drug effects; D006657:Histones; D006801:Humans; D018386:Kinetochores / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D008938:Mitosis / Q000187:drug effects; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000737:chemistry / Q000378:metabolism; D009154:Mutation / Q000235:genetics; D015739:Nocodazole / Q000494:pharmacology; D009687:Nuclear Proteins; D011485:Protein Binding; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D008941:Spindle Apparatus / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology*
C485917:CENPU protein, human; D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D006657:Histones; D046912:Multiprotein Complexes; D009687:Nuclear Proteins; D015739:Nocodazole