Bub3 interaction with Mad2, Mad3 and Cdc20 is mediated by WD40 repeats and does not require intact kinetochores

被引:156
作者
Fraschini, R
Beretta, A
Sironi, L
Musacchio, A
Lucchini, G
Piatti, S
机构
[1] Dipartimento Biotecnol & Biosci, I-20126 Milan, Italy
[2] European Inst Oncol, Dept Expt Oncol, I-20141 Milan, Italy
关键词
Bub3; Cdc20; kinetochore; mitotic checkpoint; WD40;
D O I
10.1093/emboj/20.23.6648
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The kinetochore checkpoint pathway, involving the Mad1, Mad2, Mad3, Bub1, Bub3 and Mps1 proteins, prevents anaphase entry and mitotic exit by inhibiting the anaphase promoting complex activator Cdc20 in response to monopolar attachment of sister kinetochores to spindle fibres. We show here that Cdc20, which had previously been shown to interact physically with Mad2 and Mad3, associates also with Bub3 an association is up-regulated upon checkpoint activation. Moreover, co-fractionation experiments suggest that Mad2, Mad3 and Bub3 may be concomitantly present in protein complexes with Cdc20. Formation of the Bub3-Cdc20 complex requires all kinetochore checkpoint proteins but, surprisingly, not intact kinetochores. Conversely, point mutations altering the conserved WD40 motifs of Bub3, which might be involved in the formation of a beta -propeller fold devoted to protein-protein interactions, disrupt its association with Mad2. Mad3 and Cdc20. as well as proper checkpoint response. We suggest that Bub3 could serve as a platform for interactions between kinetochore checkpoint proteins, and its association with Mad2, Mad3 and Cdc20 might be instrumental for checkpoint activation.
引用
收藏
页码:6648 / 6659
页数:12
相关论文
共 61 条
  • [1] Sister chromatid separation and chromosome re-duplication are regulated by different mechanisms in response to spindle damage
    Alexandru, G
    Zachariae, W
    Schleiffer, A
    Nasmyth, K
    [J]. EMBO JOURNAL, 1999, 18 (10) : 2707 - 2721
  • [2] A mechanism for coupling exit from mitosis to partitioning of the nucleus
    Bardin, AJ
    Visintin, R
    Amon, A
    [J]. CELL, 2000, 102 (01) : 21 - 31
  • [3] Localization of the Drosophila checkpoint control protein Bub3 to the kinetochore requires Bub1 but not Zw10 or Rod
    Basu, J
    Logarinho, E
    Herrmann, S
    Bousbaa, H
    Li, ZX
    Chan, GKT
    Yen, TJ
    Sunkel, CE
    Goldberg, ML
    [J]. CHROMOSOMA, 1998, 107 (6-7) : 376 - 385
  • [4] Anaphase spindle position is monitored by the BUB2 checkpoint
    Bloecher, A
    Venturi, GM
    Tatchell, K
    [J]. NATURE CELL BIOLOGY, 2000, 2 (08) : 556 - 558
  • [5] Complex formation between Mad1p, Bub1p and Bub3p is crucial for spindle checkpoint function
    Brady, DM
    Hardwick, KG
    [J]. CURRENT BIOLOGY, 2000, 10 (11) : 675 - 678
  • [6] The spindle checkpoint of budding yeast depends on a tight complex between the Mad1 and Mad2 proteins
    Chen, RH
    Brady, DM
    Smith, D
    Murray, AW
    Hardwick, KG
    [J]. MOLECULAR BIOLOGY OF THE CELL, 1999, 10 (08) : 2607 - 2618
  • [7] Spindle checkpoint protein Xmad1 recruits Xmad2 to unattached kinetochores
    Chen, RH
    Shevchenko, A
    Mann, M
    Murray, AW
    [J]. JOURNAL OF CELL BIOLOGY, 1998, 143 (02) : 283 - 295
  • [8] Association of spindle assembly checkpoint component XMAD2 with unattached kinetochores
    Chen, RH
    Waters, JC
    Salmon, ED
    Murray, AW
    [J]. SCIENCE, 1996, 274 (5285) : 242 - 246
  • [9] An ESP1/PDS1 complex regulates loss of sister chromatid cohesion at the metaphase to anaphase transition in yeast
    Ciosk, R
    Zachariae, W
    Michaelis, C
    Shevchenko, A
    Mann, M
    Nasmyth, K
    [J]. CELL, 1998, 93 (06) : 1067 - 1076
  • [10] CLB5 - A NOVEL B-CYCLIN FROM BUDDING YEAST WITH A ROLE IN S-PHASE
    EPSTEIN, CB
    CROSS, FR
    [J]. GENES & DEVELOPMENT, 1992, 6 (09) : 1695 - 1706