Spc24 and Stu2 Promote Spindle Integrity when DNA Replication is Stalled

Overview

Journal Mol Biol Cell

Specialties Cell Biology
Molecular Biology

Date 2007 May 18

PMID 17507656

Citations 17

Authors

Lina Ma

Jennifer McQueen

Lara Cuschieri

Jackie Vogel

Vivien Measday

Affiliations

Soon will be listed here.

Abstract

The kinetochore, a protein complex that links chromosomes to microtubules (MTs), is required to prevent spindle expansion during S phase in budding yeast, but the mechanism of how the kinetochore maintains integrity of the bipolar spindle before mitosis is not well understood. Here, we demonstrate that a mutation of Spc24, a component of the conserved Ndc80 kinetochore complex, causes lethality when cells are exposed to the DNA replication inhibitor hydroxyurea (HU) due to premature spindle expansion and segregation of incompletely replicated DNA. Overexpression of Stu1, a CLASP-related MT-associated protein or a truncated form of the XMAP215 orthologue Stu2 rescues spc24-9 HU lethality and prevents spindle expansion. Truncated Stu2 likely acts in a dominant-negative manner, because overexpression of full-length STU2 does not rescue spc24-9 HU lethality, and spindle expansion in spc24-9 HU-treated cells requires active Stu2. Stu1 and Stu2 localize to the kinetochore early in the cell cycle and Stu2 kinetochore localization depends on Spc24. We propose that mislocalization of Stu2 results in premature spindle expansion in S phase stalled spc24-9 mutants. Identifying factors that restrain spindle expansion upon inhibition of DNA replication is likely applicable to the mechanism by which spindle elongation is regulated during a normal cell cycle.

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References

Pramila T, Wu W, Miles S, Noble W, Breeden L . The Forkhead transcription factor Hcm1 regulates chromosome segregation genes and fills the S-phase gap in the transcriptional circuitry of the cell cycle. Genes Dev. 2006; 20(16):2266-78. PMC: 1553209. DOI: 10.1101/gad.1450606. View

Cuschieri L, Miller R, Vogel J . Gamma-tubulin is required for proper recruitment and assembly of Kar9-Bim1 complexes in budding yeast. Mol Biol Cell. 2006; 17(10):4420-34. PMC: 1635365. DOI: 10.1091/mbc.e06-03-0245. View

Al-Bassam J, Larsen N, Hyman A, Harrison S . Crystal structure of a TOG domain: conserved features of XMAP215/Dis1-family TOG domains and implications for tubulin binding. Structure. 2007; 15(3):355-62. DOI: 10.1016/j.str.2007.01.012. View

Zhu G, Davis T . The fork head transcription factor Hcm1p participates in the regulation of SPC110, which encodes the calmodulin-binding protein in the yeast spindle pole body. Biochim Biophys Acta. 1999; 1448(2):236-44. DOI: 10.1016/s0167-4889(98)00135-9. View

Hyland K, Kingsbury J, Koshland D, Hieter P . Ctf19p: A novel kinetochore protein in Saccharomyces cerevisiae and a potential link between the kinetochore and mitotic spindle. J Cell Biol. 1999; 145(1):15-28. PMC: 2148226. DOI: 10.1083/jcb.145.1.15. View

Adams I, Kilmartin J . Localization of core spindle pole body (SPB) components during SPB duplication in Saccharomyces cerevisiae. J Cell Biol. 1999; 145(4):809-23. PMC: 2133189. DOI: 10.1083/jcb.145.4.809. View

Krishnan V, Nirantar S, Crasta K, Cheng A, Surana U . DNA replication checkpoint prevents precocious chromosome segregation by regulating spindle behavior. Mol Cell. 2004; 16(5):687-700. DOI: 10.1016/j.molcel.2004.11.001. View

Sarin S, Ross K, Boucher L, Green Y, Tyers M, Cohen-Fix O . Uncovering novel cell cycle players through the inactivation of securin in budding yeast. Genetics. 2004; 168(3):1763-71. PMC: 1448791. DOI: 10.1534/genetics.104.029033. View

Westermann S, Avila-Sakar A, Wang H, Niederstrasser H, Wong J, Drubin D . Formation of a dynamic kinetochore- microtubule interface through assembly of the Dam1 ring complex. Mol Cell. 2005; 17(2):277-90. DOI: 10.1016/j.molcel.2004.12.019. View

10.

Miranda J, De Wulf P, Sorger P, Harrison S . The yeast DASH complex forms closed rings on microtubules. Nat Struct Mol Biol. 2005; 12(2):138-43. DOI: 10.1038/nsmb896. View

11.

Bachant J, Jessen S, Kavanaugh S, Fielding C . The yeast S phase checkpoint enables replicating chromosomes to bi-orient and restrain spindle extension during S phase distress. J Cell Biol. 2005; 168(7):999-1012. PMC: 2171834. DOI: 10.1083/jcb.200412076. View

12.

Wei R, Sorger P, Harrison S . Molecular organization of the Ndc80 complex, an essential kinetochore component. Proc Natl Acad Sci U S A. 2005; 102(15):5363-7. PMC: 555962. DOI: 10.1073/pnas.0501168102. View

13.

Inoue Y, do Carmo Avides M, Shiraki M, Deak P, Yamaguchi M, Nishimoto Y . Orbit, a novel microtubule-associated protein essential for mitosis in Drosophila melanogaster. J Cell Biol. 2000; 149(1):153-66. PMC: 2175100. DOI: 10.1083/jcb.149.1.153. View

14.

Goshima G, Yanagida M . Establishing biorientation occurs with precocious separation of the sister kinetochores, but not the arms, in the early spindle of budding yeast. Cell. 2000; 100(6):619-33. DOI: 10.1016/s0092-8674(00)80699-6. View

15.

Arino J, Saito H, Sunnerhagen P, Posas F . Rck2 kinase is a substrate for the osmotic stress-activated mitogen-activated protein kinase Hog1. Mol Cell Biol. 2000; 20(11):3887-95. PMC: 85729. DOI: 10.1128/MCB.20.11.3887-3895.2000. View

16.

He X, Asthana S, Sorger P . Transient sister chromatid separation and elastic deformation of chromosomes during mitosis in budding yeast. Cell. 2000; 101(7):763-75. DOI: 10.1016/s0092-8674(00)80888-0. View

17.

Cheeseman I, Muller-Reichert T, Drubin D, Barnes G . Mitotic spindle integrity and kinetochore function linked by the Duo1p/Dam1p complex. J Cell Biol. 2001; 152(1):197-212. PMC: 2193660. DOI: 10.1083/jcb.152.1.197. View

18.

Janke C, Ortiz J, Lechner J, Shevchenko A, Magiera M, Schramm C . The budding yeast proteins Spc24p and Spc25p interact with Ndc80p and Nuf2p at the kinetochore and are important for kinetochore clustering and checkpoint control. EMBO J. 2001; 20(4):777-91. PMC: 145434. DOI: 10.1093/emboj/20.4.777. View

19.

Canman C . Replication checkpoint: preventing mitotic catastrophe. Curr Biol. 2001; 11(4):R121-4. DOI: 10.1016/s0960-9822(01)00057-4. View

20.

PEARSON C, Maddox P, Salmon E, Bloom K . Budding yeast chromosome structure and dynamics during mitosis. J Cell Biol. 2001; 152(6):1255-66. PMC: 2199205. DOI: 10.1083/jcb.152.6.1255. View