INCENP is required for proper targeting of Survivin to the centromeres and the anaphase spindle during mitosis.
Three lines of investigation have suggested that interactions between Survivin and the chromosomal passenger proteins INCENP and Aurora-B kinase may be important for mitotic progression. First, interference with the function of Survivin/BIR1, INCENP, or Aurora-B kinase leads to similar defects in mitosis and cytokinesis [1-7] (see [8] for review). Second, INCENP and Aurora-B exist in a complex in Xenopus eggs [9] and in mammalian cultured cells [7]. Third, interference with Survivin or INCENP function causes Aurora-B kinase to be mislocalized in mitosis in both C. elegans and vertebrates [5, 7, 9]. Here, we provide evidence that Survivin, Aurora-B, and INCENP interact physically and functionally. Direct visualization of Survivin-GFP in mitotic cells reveals that it localizes identically to INCENP and Aurora-B. Survivin binds directly to both Aurora-B and INCENP in yeast two-hybrid and in vitro pull-down assays. The in vitro interaction between Survivin and Aurora-B is extraordinarily stable in that it resists 3 M NaCl. Finally, Survivin and INCENP interact functionally in vivo; in cells in which INCENP localization is disrupted, Survivin adheres to the chromosomes and no longer concentrates at the centromeres or transfers to the anaphase spindle midzone. Our data provide the first biochemical evidence that Survivin can interact directly with members of the chromosomal passenger complex.
Wheatley
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SP
Chromosome Structure Group, Institute of Cell and Molecular Biology, Swann Building, University of Edinburgh, King's Buildings, Mayfield Road, EH9 3JR, Edinburgh, United Kingdom.
Carvalho
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A
Vagnarelli
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P
Earnshaw
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WC
D000705:Anaphase / Q000502:physiology*; D000818:Animals; D064107:Aurora Kinase B; D064127:Aurora Kinases; D002352:Carrier Proteins / Q000378:metabolism; D002451:Cell Compartmentation; D002503:Centromere / Q000378:metabolism* / Q000648:ultrastructure; D002645:Chickens; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009154:Mutation; D009363:Neoplasm Proteins; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000302:isolation & purification / Q000378:metabolism; D021381:Protein Transport; D008941:Spindle Apparatus / Q000378:metabolism* / Q000648:ultrastructure; D000077022:Survivin; D014407:Tumor Cells, Cultured; D020798:Two-Hybrid System Techniques
C107489:BIRC5 protein, human; D002352:Carrier Proteins; D002868:Chromosomal Proteins, Non-Histone; C083767:INCENP protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D000077022:Survivin; C576921:AURKB protein, human; D064107:Aurora Kinase B; D064127:Aurora Kinases; D017346:Protein Serine-Threonine Kinases