Home LiteratureArticle Details
PMID: 15930123 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Tension-dependent regulation of microtubule dynamics at kinetochores can explain metaphase congression in yeast.

Molecular biology of the cell ·Vol. 16 ·No. 8 ·2005-08-00 ·Pages 3764-75

Gardner MK, Pearson CG, Sprague BL, Zarzar TR, Bloom K, Salmon ED, Odde DJ

Abstract

During metaphase in budding yeast mitosis, sister kinetochores are tethered to opposite poles and separated, stretching their intervening chromatin, by singly attached kinetochore microtubules (kMTs). Kinetochore movements are coupled to single microtubule plus-end polymerization/depolymerization at kinetochore attachment sites. Here, we use computer modeling to test possible mechanisms controlling chromosome alignment during yeast metaphase by simulating experiments that determine the 1) mean positions of kinetochore Cse4-GFP, 2) extent of oscillation of kinetochores during metaphase as measured by fluorescence recovery after photobleaching (FRAP) of kinetochore Cse4-GFP, 3) dynamics of kMTs as measured by FRAP of GFP-tubulin, and 4) mean positions of unreplicated chromosome kinetochores that lack pulling forces from a sister kinetochore. We rule out a number of possible models and find the best fit between theory and experiment when it is assumed that kinetochores sense both a spatial gradient that suppresses kMT catastrophe near the poles and attachment site tension that promotes kMT rescue at higher amounts of chromatin stretch.

MeSH Terms
Cell Cycle Proteins/genetics,metabolism Chromatin/metabolism Chromosomal Proteins, Non-Histone DNA-Binding Proteins/metabolism Kinetochores/metabolism Metaphase Microtubules/metabolism Models, Biological Mutation/genetics Recombinant Proteins/genetics,metabolism Saccharomyces cerevisiae/cytology,metabolism Saccharomyces cerevisiae Proteins/genetics,metabolism Spectrometry, Fluorescence
Chemicals
CDC6 protein, S cerevisiae CSE4 protein, S cerevisiae Cell Cycle Proteins Chromatin Chromosomal Proteins, Non-Histone DNA-Binding Proteins Recombinant Proteins Saccharomyces cerevisiae Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Gardner Melissa K
Department of Biomedical Engineering, University of Minnesota, Minneapolis, MN 55455, USA.
Pearson Chad G
Sprague Brian L
Zarzar Ted R
Bloom Kerry
Salmon E D
Odde David J
References (53)
53 references, click to expand
  1. Mechanical disruption of individual nucleosomes reveals a reversible multistage release of DNA.
    Proc Natl Acad Sci U S A. 2002 Feb 19;99(4):1960-5 PMID: 11854495
  2. Transient sister chromatid separation and elastic deformation of chromosomes during mitosis in budding yeast.
    Cell. 2000 Jun 23;101(7):763-75 PMID: 10892747
  3. Simple centromere, complex kinetochore: linking spindle microtubules and centromeric DNA in budding yeast.
    J Cell Biol. 2002 Apr 15;157(2):199-203 PMID: 11956223
  4. Estimates of lateral and longitudinal bond energies within the microtubule lattice.
    Proc Natl Acad Sci U S A. 2002 Apr 30;99(9):6035-40 PMID: 11983898
  5. Segregating sister genomes: the molecular biology of chromosome separation.
    Science. 2002 Jul 26;297(5581):559-65 PMID: 12142526
  6. beta-Tubulin C354 mutations that severely decrease microtubule dynamics do not prevent nuclear migration in yeast.
    Mol Biol Cell. 2002 Aug;13(8):2919-32 PMID: 12181356
  7. Attachment and tension in the spindle assembly checkpoint.
    J Cell Sci. 2002 Sep 15;115(Pt 18):3547-55 PMID: 12186941
  8. Centromeres and kinetochores: from epigenetics to mitotic checkpoint signaling.
    Cell. 2003 Feb 21;112(4):407-21 PMID: 12600307
  9. Cell division.
    Nature. 2003 Apr 17;422(6933):746-52 PMID: 12700768
  10. Dynamics and mechanics of the microtubule plus end.
    Nature. 2003 Apr 17;422(6933):753-8 PMID: 12700769
  11. Stu2p, the budding yeast member of the conserved Dis1/XMAP215 family of microtubule-associated proteins is a plus end-binding microtubule destabilizer.
    J Cell Biol. 2003 Apr 28;161(2):359-69 PMID: 12719475
  12. Mechanisms of microtubule-based kinetochore positioning in the yeast metaphase spindle.
    Biophys J. 2003 Jun;84(6):3529-46 PMID: 12770865
  13. Captivating capture: how microtubules attach to kinetochores.
    Curr Biol. 2003 May 27;13(11):R449-60 PMID: 12781157
  14. 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
  15. Yeast kinetochores do not stabilize Stu2p-dependent spindle microtubule dynamics.
    Mol Biol Cell. 2003 Oct;14(10):4181-95 PMID: 14517328
  16. Tension between two kinetochores suffices for their bi-orientation on the mitotic spindle.
    Nature. 2004 Mar 4;428(6978):93-7 PMID: 14961024
  17. Microtubule plus-end dynamics in Xenopus egg extract spindles.
    Mol Biol Cell. 2004 Apr;15(4):1776-84 PMID: 14767058
  18. Electron-microscopic study of the spindle and chromosome movement in the yeast Saccharomyces cerevisiae.
    J Cell Sci. 1976 Nov;22(2):219-42 PMID: 794073
  19. The forces that move chromosomes in mitosis.
    Annu Rev Biophys Biophys Chem. 1988;17:431-49 PMID: 3293594
  20. 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
  21. Motile kinetochores and polar ejection forces dictate chromosome position on the vertebrate mitotic spindle.
    J Cell Biol. 1994 Feb;124(3):223-33 PMID: 8294508
  22. Elements of error correction in mitosis: microtubule capture, release, and tension.
    J Cell Biol. 1994 Sep;126(5):1241-53 PMID: 8063861
  23. Three-dimensional ultrastructural analysis of the Saccharomyces cerevisiae mitotic spindle.
    J Cell Biol. 1995 Jun;129(6):1601-15 PMID: 7790357
  24. Kinetochore motility after severing between sister centromeres using laser microsurgery: evidence that kinetochore directional instability and position is regulated by tension.
    J Cell Sci. 1995 Jul;108 ( Pt 7):2537-48 PMID: 7593295
  25. Force generation by microtubule assembly/disassembly in mitosis and related movements.
    Mol Biol Cell. 1995 Dec;6(12):1619-40 PMID: 8590794
  26. Evidence that a single monolayer tubulin-GTP cap is both necessary and sufficient to stabilize microtubules.
    Mol Biol Cell. 1996 Apr;7(4):663-75 PMID: 8730106
  27. Centromere position in budding yeast: evidence for anaphase A.
    Mol Biol Cell. 1997 Jun;8(6):957-72 PMID: 9201708
  28. Microtubules orient the mitotic spindle in yeast through dynein-dependent interactions with the cell cortex.
    J Cell Biol. 1997 Aug 11;138(3):629-41 PMID: 9245791
  29. 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
  30. Astral microtubule dynamics in yeast: a microtubule-based searching mechanism for spindle orientation and nuclear migration into the bud.
    J Cell Biol. 1997 Nov 17;139(4):985-94 PMID: 9362516
  31. Cell cycle: checkpoint proteins and kinetochores.
    Curr Biol. 1997 Oct 1;7(10):R613-6 PMID: 9368739
  32. The vertebrate cell kinetochore and its roles during mitosis.
    Trends Cell Biol. 1998 Aug;8(8):310-8 PMID: 9704407
  33. The polarity and dynamics of microtubule assembly in the budding yeast Saccharomyces cerevisiae.
    Nat Cell Biol. 2000 Jan;2(1):36-41 PMID: 10620805
  34. Rings around kinetochore microtubules in yeast.
    Nat Struct Mol Biol. 2005 Mar;12(3):210-2 PMID: 15744320
  35. Cohesin ensures bipolar attachment of microtubules to sister centromeres and resists their precocious separation.
    Nat Cell Biol. 2000 Aug;2(8):492-9 PMID: 10934469
  36. The N terminus of the centromere H3-like protein Cse4p performs an essential function distinct from that of the histone fold domain.
    Mol Cell Biol. 2000 Sep;20(18):7037-48 PMID: 10958698
  37. Budding yeast chromosome structure and dynamics during mitosis.
    J Cell Biol. 2001 Mar 19;152(6):1255-66 PMID: 11257125
  38. Molecular analysis of kinetochore-microtubule attachment in budding yeast.
    Cell. 2001 Jul 27;106(2):195-206 PMID: 11511347
  39. Control of microtubule dynamics by Stu2p is essential for spindle orientation and metaphase chromosome alignment in yeast.
    Mol Biol Cell. 2001 Sep;12(9):2870-80 PMID: 11553724
  40. Time course analysis of precocious separation of sister centromeres in budding yeast: continuously separated or frequently reassociated?
    Genes Cells. 2001 Sep;6(9):765-73 PMID: 11554923
  41. Lack of tension at kinetochores activates the spindle checkpoint in budding yeast.
    Curr Biol. 2001 Sep 18;11(18):1462-7 PMID: 11566107
  42. The budding yeast protein kinase Ipl1/Aurora allows the absence of tension to activate the spindle checkpoint.
    Genes Dev. 2001 Dec 1;15(23):3118-29 PMID: 11731476
  43. Checkpoint signals in grasshopper meiosis are sensitive to microtubule attachment, but tension is still essential.
    J Cell Sci. 2001 Dec;114(Pt 23):4173-83 PMID: 11739650
  44. Cse4p is a component of the core centromere of Saccharomyces cerevisiae.
    Cell. 1998 Sep 4;94(5):607-13 PMID: 9741625
  45. Yeast Bim1p promotes the G1-specific dynamics of microtubules.
    J Cell Biol. 1999 May 31;145(5):993-1007 PMID: 10352017
  46. High-voltage electron tomography of spindle pole bodies and early mitotic spindles in the yeast Saccharomyces cerevisiae.
    Mol Biol Cell. 1999 Jun;10(6):2017-31 PMID: 10359612
  47. Spatial gradients of cellular phospho-proteins.
    FEBS Lett. 1999 Sep 3;457(3):452-4 PMID: 10471827
  48. Stable kinetochore-microtubule attachment constrains centromere positioning in metaphase.
    Curr Biol. 2004 Nov 9;14(21):1962-7 PMID: 15530400
  49. DNA replication checkpoint prevents precocious chromosome segregation by regulating spindle behavior.
    Mol Cell. 2004 Dec 3;16(5):687-700 PMID: 15574325
  50. Anaphase spindle mechanics prevent mis-segregation of merotelically oriented chromosomes.
    Curr Biol. 2004 Dec 14;14(23):2149-55 PMID: 15589159
  51. Formation of a dynamic kinetochore- microtubule interface through assembly of the Dam1 ring complex.
    Mol Cell. 2005 Jan 21;17(2):277-90 PMID: 15664196
  52. The yeast DASH complex forms closed rings on microtubules.
    Nat Struct Mol Biol. 2005 Feb;12(2):138-43 PMID: 15640796
  53. Visualization of a Ran-GTP gradient in interphase and mitotic Xenopus egg extracts.
    Science. 2002 Mar 29;295(5564):2452-6 PMID: 11923538
Article Info
Journal
Molecular biology of the cell
Abbr.
Mol Biol Cell
ISSN
1059-1524
DOI
10.1091/mbc.e05-04-0275
Published
2005-08-00
Epub
2005-00-01
Pages
3764-75
Language
English
Region
United States
NLM ID
9201390
PMCID
PMC1182314
Subset
IM
Grants
NIGMS NIH HHS · GM-24364 · United States
NIGMS NIH HHS · R01 GM032238 · United States
NIGMS NIH HHS · GM-32238 · United States
NIGMS NIH HHS · R37 GM032238 · United States
NIGMS NIH HHS · R37 GM024364 · United States
NIGMS NIH HHS · R01 GM024364 · United States
External Links
PubMed source DOI: 10.1091/mbc.e05-04-0275 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2