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PMID: 14699064 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Depletion of centromeric MCAK leads to chromosome congression and segregation defects due to improper kinetochore attachments.

Molecular biology of the cell ·Vol. 15 ·No. 3 ·2004-03-00 ·Pages 1146-59

Kline-Smith SL, Khodjakov A, Hergert P, Walczak CE

Abstract

The complex behavior of chromosomes during mitosis is accomplished by precise binding and highly regulated polymerization dynamics of kinetochore microtubules. Previous studies have implicated Kin Is, unique kinesins that depolymerize microtubules, in regulating chromosome positioning. We have characterized the immunofluorescence localization of centromere-bound MCAK and found that MCAK localized to inner kinetochores during prophase but was predominantly centromeric by metaphase. Interestingly, MCAK accumulated at leading kinetochores during congression but not during segregation. We tested the consequences of MCAK disruption by injecting a centromere dominant-negative protein into prophase cells. Depletion of centromeric MCAK led to reduced centromere stretch, delayed chromosome congression, alignment defects, and severe missegregation of chromosomes. Rates of chromosome movement were unchanged, suggesting that the primary role of MCAK is not to move chromosomes. Furthermore, we found that disruption of MCAK leads to multiple kinetochore-microtubule attachment defects, including merotelic, syntelic, and combined merotelic-syntelic attachments. These findings reveal an essential role for Kin Is in prevention and/or correction of improper kinetochore-microtubule attachments.

MeSH Terms
Animals Cell Cycle/physiology Cells, Cultured Chromosome Segregation/physiology Chromosomes/metabolism Humans Kinesins/metabolism Kinetochores/metabolism Microscopy, Electron Microtubules/metabolism Spindle Apparatus/metabolism Xenopus/metabolism
Chemicals
KIF2C protein, human Kinesins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kline-Smith Susan L
Departments of Anatomy and Cell Biology, Indiana University Medical Sciences Program, Bloomington, Indiana 47405, USA.
Khodjakov Alexey
Hergert Polla
Walczak Claire E
References (52)
52 references, click to expand
  1. The small molecule Hesperadin reveals a role for Aurora B in correcting kinetochore-microtubule attachment and in maintaining the spindle assembly checkpoint.
    J Cell Biol. 2003 Apr 28;161(2):281-94 PMID: 12707311
  2. The conserved protein kinase Ipl1 regulates microtubule binding to kinetochores in budding yeast.
    Genes Dev. 1999 Mar 1;13(5):532-44 PMID: 10072382
  3. Unconventional motoring: an overview of the Kin C and Kin I kinesins.
    Traffic. 2003 Jun;4(6):367-75 PMID: 12753646
  4. Direct observation of microtubule dynamics at kinetochores in Xenopus extract spindles: implications for spindle mechanics.
    J Cell Biol. 2003 Aug 4;162(3):377-82 PMID: 12900391
  5. An inner centromere protein that stimulates the microtubule depolymerizing activity of a KinI kinesin.
    Dev Cell. 2003 Aug;5(2):309-21 PMID: 12919681
  6. Merotelic kinetochore orientation occurs frequently during early mitosis in mammalian tissue cells and error correction is achieved by two different mechanisms.
    J Cell Sci. 2003 Oct 15;116(Pt 20):4213-25 PMID: 12953065
  7. The roles of microtubule-based motor proteins in mitosis: comprehensive RNAi analysis in the Drosophila S2 cell line.
    J Cell Biol. 2003 Sep 15;162(6):1003-16 PMID: 12975346
  8. Light and electron microscopy of rat kangaroo cells in mitosis. II. Kinetochore structure and function.
    Chromosoma. 1973;41(2):195-220 PMID: 4571311
  9. Nucleated assembly of mitotic microtubules in living PTK2 cells after release from nocodazole treatment.
    Cell Motil. 1981;1(4):469-83 PMID: 7348606
  10. The formation, structure, and composition of the mammalian kinetochore and kinetochore fiber.
    Int Rev Cytol. 1982;79:1-58 PMID: 6185450
  11. Stability of microtubule attachment to metaphase kinetochores in PtK1 cells.
    J Cell Sci. 1990 May;96 ( Pt 1):9-15 PMID: 2197288
  12. Chromosome fiber dynamics and congression oscillations in metaphase PtK2 cells at 23 degrees C.
    Cell Motil Cytoskeleton. 1991;18(2):131-42 PMID: 2013109
  13. Directional instability of kinetochore motility during chromosome congression and segregation in mitotic newt lung cells: a push-pull mechanism.
    J Cell Biol. 1993 Aug;122(4):859-75 PMID: 8349735
  14. Differential expression of a phosphoepitope at the kinetochores of moving chromosomes.
    J Cell Biol. 1993 Sep;122(6):1311-21 PMID: 7690762
  15. Active zones on motor nerve terminals contain alpha 3beta 1 integrin.
    J Neurosci. 2000 Jul 1;20(13):4912-21 PMID: 10864949
  16. Probing spindle assembly mechanisms with monastrol, a small molecule inhibitor of the mitotic kinesin, Eg5.
    J Cell Biol. 2000 Sep 4;150(5):975-88 PMID: 10973989
  17. Tension on chromosomes increases the number of kinetochore microtubules but only within limits.
    J Cell Sci. 2000 Nov;113 Pt 21:3815-23 PMID: 11034909
  18. Chromosome movement in mitosis requires microtubule anchorage at spindle poles.
    J Cell Biol. 2001 Feb 5;152(3):425-34 PMID: 11157972
  19. Merotelic kinetochore orientation is a major mechanism of aneuploidy in mitotic mammalian tissue cells.
    J Cell Biol. 2001 Apr 30;153(3):517-27 PMID: 11331303
  20. Microtubule-dependent changes in assembly of microtubule motor proteins and mitotic spindle checkpoint proteins at PtK1 kinetochores.
    Mol Biol Cell. 2001 Jul;12(7):1995-2009 PMID: 11451998
  21. Merotelic kinetochore orientation versus chromosome mono-orientation in the origin of lagging chromosomes in human primary cells.
    J Cell Sci. 2002 Feb 1;115(Pt 3):507-15 PMID: 11861758
  22. Kinesins klp5(+) and klp6(+) are required for normal chromosome movement in mitosis.
    J Cell Sci. 2002 Mar 1;115(Pt 5):931-40 PMID: 11870212
  23. Two kinesin-like Kin I family proteins in fission yeast regulate the establishment of metaphase and the onset of anaphase A.
    Curr Biol. 2002 Apr 16;12(8):610-21 PMID: 11967147
  24. Bi-orienting chromosomes on the mitotic spindle.
    Curr Opin Cell Biol. 2002 Jun;14(3):365-71 PMID: 12067660
  25. Chromosome dynamics: new light on Aurora B kinase function.
    Curr Biol. 2002 Jul 9;12(13):R458-60 PMID: 12121637
  26. The microtubule-destabilizing kinesin XKCM1 regulates microtubule dynamic instability in cells.
    Mol Biol Cell. 2002 Aug;13(8):2718-31 PMID: 12181341
  27. Attachment and tension in the spindle assembly checkpoint.
    J Cell Sci. 2002 Sep 15;115(Pt 18):3547-55 PMID: 12186941
  28. The spindle checkpoint: structural insights into dynamic signalling.
    Nat Rev Mol Cell Biol. 2002 Oct;3(10):731-41 PMID: 12360190
  29. Chromosome-microtubule interactions during mitosis.
    Annu Rev Cell Dev Biol. 2002;18:193-219 PMID: 12142285
  30. Mad2 and BubR1 function in a single checkpoint pathway that responds to a loss of tension.
    Mol Biol Cell. 2002 Oct;13(10):3706-19 PMID: 12388768
  31. The microtubule-destabilizing kinesin XKCM1 is required for chromosome positioning during spindle assembly.
    Curr Biol. 2002 Oct 29;12(21):1885-9 PMID: 12419191
  32. The kinesin-related protein MCAK is a microtubule depolymerase that forms an ATP-hydrolyzing complex at microtubule ends.
    Mol Cell. 2003 Feb;11(2):445-57 PMID: 12620232
  33. Microtubule assembly and kinetochore directional instability in vertebrate monopolar spindles: implications for the mechanism of chromosome congression.
    J Cell Sci. 1994 Jan;107 ( Pt 1):285-97 PMID: 8175915
  34. High resolution multimode digital imaging system for mitosis studies in vivo and in vitro.
    Biol Bull. 1994 Oct;187(2):231-2 PMID: 7811791
  35. Identification and partial characterization of mitotic centromere-associated kinesin, a kinesin-related protein that associates with centromeres during mitosis.
    J Cell Biol. 1995 Jan;128(1-2):95-104 PMID: 7822426
  36. Kinetochore chemistry is sensitive to tension and may link mitotic forces to a cell cycle checkpoint.
    J Cell Biol. 1995 Aug;130(4):929-39 PMID: 7642708
  37. XKCM1: a Xenopus kinesin-related protein that regulates microtubule dynamics during mitotic spindle assembly.
    Cell. 1996 Jan 12;84(1):37-47 PMID: 8548824
  38. Force generation by microtubule assembly/disassembly in mitosis and related movements.
    Mol Biol Cell. 1995 Dec;6(12):1619-40 PMID: 8590794
  39. Sterecon--three-dimensional reconstructions from stereoscopic contouring.
    J Struct Biol. 1996 Jan-Feb;116(1):93-8 PMID: 8742729
  40. Modulation of CENP-E organization at kinetochores by spindle microtubule attachment.
    Cell Motil Cytoskeleton. 1996;35(2):121-33 PMID: 8894282
  41. The kinetochore microtubule minus-end disassembly associated with poleward flux produces a force that can do work.
    Mol Biol Cell. 1996 Oct;7(10):1547-58 PMID: 8898361
  42. How cells get the right chromosomes.
    Science. 1997 Jan 31;275(5300):632-7 PMID: 9005842
  43. Oscillating mitotic newt lung cell kinetochores are, on average, under tension and rarely push.
    J Cell Sci. 1996 Dec;109 ( Pt 12):2823-31 PMID: 9013330
  44. Chromosome fragments possessing only one kinetochore can congress to the spindle equator.
    J Cell Biol. 1997 Jan 27;136(2):229-40 PMID: 9015296
  45. Kinetochore fiber maturation in PtK1 cells and its implications for the mechanisms of chromosome congression and anaphase onset.
    J Cell Biol. 1997 Jun 30;137(7):1567-80 PMID: 9199171
  46. Micromanipulation of chromosomes in mitotic vertebrate tissue cells: tension controls the state of kinetochore movement.
    Exp Cell Res. 1997 Sep 15;235(2):314-24 PMID: 9299155
  47. Mitotic centromere-associated kinesin is important for anaphase chromosome segregation.
    J Cell Biol. 1998 Aug 10;142(3):787-801 PMID: 9700166
  48. The vertebrate cell kinetochore and its roles during mitosis.
    Trends Cell Biol. 1998 Aug;8(8):310-8 PMID: 9704407
  49. Correlative light and electron microscopy of mitotic cells in monolayer cultures.
    Methods Cell Biol. 1999;61:297-315 PMID: 9891321
  50. The hBUB1 and hBUBR1 kinases sequentially assemble onto kinetochores during prophase with hBUBR1 concentrating at the kinetochore plates in mitosis.
    Chromosoma. 1998 Dec;107(6-7):386-96 PMID: 9914370
  51. Kin I kinesins are microtubule-destabilizing enzymes.
    Cell. 1999 Jan 8;96(1):69-78 PMID: 9989498
  52. Aurora B couples chromosome alignment with anaphase by targeting BubR1, Mad2, and Cenp-E to kinetochores.
    J Cell Biol. 2003 Apr 28;161(2):267-80 PMID: 12719470
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e03-08-0581
Published
2004-03-00
Epub
2003-00-29
Pages
1146-59
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC363095
Subset
IM
Grants
NIGMS NIH HHS · R01 GM059363 · United States
NIGMS NIH HHS · R01 GM059363-05 · United States
NIGMS NIH HHS · R01 GM059618 · United States
External Links
PubMed source DOI: 10.1091/mbc.e03-08-0581 PMC Full Text View journal details
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