Loss of Function of the Cik1/Kar3 Motor Complex Results in Chromosomes with Syntelic Attachment That Are Sensed by the Tension Checkpoint

English version České info

The attachment of sister kinetochores by microtubules emanating from opposite spindle poles establishes chromosome bipolar attachment, which generates tension on chromosomes and is essential for sister-chromatid segregation. Syntelic attachment occurs when both sister kinetochores are attached by microtubules from the same spindle pole and this attachment is unable to generate tension on chromosomes, but a reliable method to induce syntelic attachments is not available in budding yeast. The spindle checkpoint can sense the lack of tension on chromosomes as well as detached kinetochores to prevent anaphase onset. In budding yeast Saccharomyces cerevisiae, tension checkpoint proteins Aurora/Ipl1 kinase and centromere-localized Sgo1 are required to sense the absence of tension but are dispensable for the checkpoint response to detached kinetochores. We have found that the loss of function of a motor protein complex Cik1/Kar3 in budding yeast leads to syntelic attachments. Inactivation of either the spindle or tension checkpoint enables premature anaphase entry in cells with dysfunctional Cik1/Kar3, resulting in co-segregation of sister chromatids. Moreover, the abolished Kar3-kinetochore interaction in cik1 mutants suggests that the Cik1/Kar3 complex mediates chromosome movement along microtubules, which could facilitate bipolar attachment. Therefore, we can induce syntelic attachments in budding yeast by inactivating the Cik1/Kar3 complex, and this approach will be very useful to study the checkpoint response to syntelic attachments.

Vyšlo v časopise: Loss of Function of the Cik1/Kar3 Motor Complex Results in Chromosomes with Syntelic Attachment That Are Sensed by the Tension Checkpoint. PLoS Genet 8(2): e32767. doi:10.1371/journal.pgen.1002492
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1002492

Souhrn

Zdroje

1. PinskyBABigginsS 2005 The spindle checkpoint: tension versus attachment. Trends Cell Biol 15 486 493

2. LiYBachantJAlcasabasAAWangYQinJ 2002 The mitotic spindle is required for loading of the DASH complex onto the kinetochore. Genes Dev 16 183 197

3. WestermannSAvila-SakarAWangHWNiederstrasserHWongJ 2005 Formation of a dynamic kinetochore- microtubule interface through assembly of the Dam1 ring complex. Mol Cell 17 277 290

4. WestermannSWangHWAvila-SakarADrubinDGNogalesE 2006 The Dam1 kinetochore ring complex moves processively on depolymerizing microtubule ends. Nature 440 565 569

5. JankeCOrtizJTanakaTULechnerJSchiebelE 2002 Four new subunits of the Dam1-Duo1 complex reveal novel functions in sister kinetochore biorientation. EMBO J 21 181 193

6. WestermannSDrubinDGBarnesG 2007 Structures and Functions of Yeast Kinetochore Complexes. Annu Rev Biochem

7. De WulfPMcAinshADSorgerPK 2003 Hierarchical assembly of the budding yeast kinetochore from multiple subcomplexes. Genes Dev 17 2902 2921

8. HoytMATotisLRobertsBT 1991 S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function. Cell 66 507 517

9. LiRMurrayAW 1991 Feedback control of mitosis in budding yeast. Cell 66 519 531

10. HardwickKGWeissELucaFCWineyMMurrayAW 1996 Activation of the budding yeast spindle assembly checkpoint without mitotic spindle disruption. Science 273 953 956

11. WangYBurkeDJ 1995 Checkpoint genes required to delay cell division in response to nocodazole respond to impaired kinetochore function in the yeast Saccharomyces cerevisiae. Mol Cell Biol 15 6838 6844

12. HardwickKGJohnstonRCSmithDLMurrayAW 2000 MAD3 encodes a novel component of the spindle checkpoint which interacts with Bub3p, Cdc20p, and Mad2p. J Cell Biol 148 871 882

13. ChenRH 2002 BubR1 is essential for kinetochore localization of other spindle checkpoint proteins and its phosphorylation requires Mad1. J Cell Biol 158 487 496

14. Cohen-FixOPetersJMKirschnerMWKoshlandD 1996 Anaphase initiation in Saccharomyces cerevisiae is controlled by the APC-dependent degradation of the anaphase inhibitor Pds1p. Genes Dev 10 3081 3093

15. CioskRZachariaeWMichaelisCShevchenkoAMannM 1998 An ESP1/PDS1 complex regulates loss of sister chromatid cohesion at the metaphase to anaphase transition in yeast. Cell 93 1067 1076

16. UhlmannFLottspeichFNasmythK 1999 Sister-chromatid separation at anaphase onset is promoted by cleavage of the cohesin subunit Scc1. Nature 400 37 42

17. LiXNicklasRB 1995 Mitotic forces control a cell-cycle checkpoint. Nature 373 630 632

18. KeatingPRachidiNTanakaTUStarkMJ 2009 Ipl1-dependent phosphorylation of Dam1 is reduced by tension applied on kinetochores. J Cell Sci 122 4375 4382

19. BigginsSMurrayAW 2001 The budding yeast protein kinase Ipl1/Aurora allows the absence of tension to activate the spindle checkpoint. Genes Dev 15 3118 3129

20. IndjeianVBSternBMMurrayAW 2005 The centromeric protein Sgo1 is required to sense lack of tension on mitotic chromosomes. Science 307 130 133

21. TanakaTURachidiNJankeCPereiraGGalovaM 2002 Evidence that the Ipl1-Sli15 (Aurora kinase-INCENP) complex promotes chromosome bi-orientation by altering kinetochore-spindle pole connections. Cell 108 317 329

22. PinskyBAKungCShokatKMBigginsS 2006 The Ipl1-Aurora protein kinase activates the spindle checkpoint by creating unattached kinetochores. Nat Cell Biol 8 78 83

23. MeluhPBRoseMD 1990 KAR3, a kinesin-related gene required for yeast nuclear fusion. Cell 60 1029 1041

24. EndowSAKangSJSatterwhiteLLRoseMDSkeenVP 1994 Yeast Kar3 is a minus-end microtubule motor protein that destabilizes microtubules preferentially at the minus ends. EMBO J 13 2708 2713

25. GardnerMKHaaseJMythreyeKMolkJNAndersonM 2008 The microtubule-based motor Kar3 and plus end-binding protein Bim1 provide structural support for the anaphase spindle. J Cell Biol 180 91 100

26. TanakaKMukaeNDewarHvan BreugelMJamesEK 2005 Molecular mechanisms of kinetochore capture by spindle microtubules. Nature 434 987 994

27. TanakaKKitamuraEKitamuraYTanakaTU 2007 Molecular mechanisms of microtubule-dependent kinetochore transport toward spindle poles. J Cell Biol 178 269 281

28. PageBDSnyderM 1992 CIK1: a developmentally regulated spindle pole body-associated protein important for microtubule functions in Saccharomyces cerevisiae. Genes Dev 6 1414 1429

29. LiuHJinFLiangFTianXWangY 2011 The Cik1/Kar3 motor complex is required for the proper kinetochore-microtubule interaction after stressful DNA replication. Genetics 187 397 407

30. DanielJAKeyesBENgYPFreemanCOBurkeDJ 2006 Diverse functions of spindle assembly checkpoint genes in Saccharomyces cerevisiae. Genetics 172 53 65

31. TongAHLesageGBaderGDDingHXuH 2004 Global mapping of the yeast genetic interaction network. Science 303 808 813

32. BarrettJGManningBDSnyderM 2000 The Kar3p kinesin-related protein forms a novel heterodimeric structure with its associated protein Cik1p. Mol Biol Cell 11 2373 2385

33. ManningBDBarrettJGWallaceJAGranokHSnyderM 1999 Differential regulation of the Kar3p kinesin-related protein by two associated proteins, Cik1p and Vik1p. J Cell Biol 144 1219 1233

34. SproulLRAndersonDJMackeyATSaundersWSGilbertSP 2005 Cik1 targets the minus-end kinesin depolymerase kar3 to microtubule plus ends. Curr Biol 15 1420 1427

35. KotwaliwaleCVFreiSBSternBMBigginsS 2007 A pathway containing the Ipl1/aurora protein kinase and the spindle midzone protein Ase1 regulates yeast spindle assembly. Dev Cell 13 433 445

36. MakrantoniVStarkMJ 2009 Efficient chromosome biorientation and the tension checkpoint in Saccharomyces cerevisiae both require Bir1. Mol Cell Biol 29 4552 4562

37. GardnerMKBouckDCPaliulisLVMeehlJBO'TooleET 2008 Chromosome congression by Kinesin-5 motor-mediated disassembly of longer kinetochore microtubules. Cell 135 894 906

38. HeXAsthanaSSorgerPK 2000 Transient sister chromatid separation and elastic deformation of chromosomes during mitosis in budding yeast. Cell 101 763 775

39. GoshimaGYanagidaM 2000 Establishing biorientation occurs with precocious separation of the sister kinetochores, but not the arms, in the early spindle of budding yeast. Cell 100 619 633

40. BachantJAlcasabasABlatYKlecknerNElledgeSJ 2002 The SUMO-1 isopeptidase Smt4 is linked to centromeric cohesion through SUMO-1 modification of DNA topoisomerase II. Mol Cell 9 1169 1182

41. TangXWangY 2006 Pds1/Esp1-dependent and -independent sister chromatid separation in mutants defective for protein phosphatase 2A. Proc Natl Acad Sci U S A 103 16290 16295

42. LiangFWangY 2007 DNA damage checkpoints inhibit mitotic exit by two different mechanisms. Mol Cell Biol 27 5067 5078

43. TytellJDSorgerPK 2006 Analysis of kinesin motor function at budding yeast kinetochores. J Cell Biol 172 861 874

44. GuacciVKoshlandDStrunnikovA 1997 A direct link between sister chromatid cohesion and chromosome condensation revealed through the analysis of MCD1 in S. cerevisiae. Cell 91 47 57

45. NgTMWaplesWGLavoieBDBigginsS 2009 Pericentromeric sister chromatid cohesion promotes kinetochore biorientation. Mol Biol Cell 20 3818 3827

46. CottinghamFRGheberLMillerDLHoytMA 1999 Novel roles for Saccharomyces cerevisiae mitotic spindle motors. J Cell Biol 147 335 350

47. NewmanJRWolfEKimPS 2000 A computationally directed screen identifying interacting coiled coils from Saccharomyces cerevisiae. Proc Natl Acad Sci U S A 97 13203 13208

48. WongJNakajimaYWestermannSShangCKangJS 2007 A protein interaction map of the mitotic spindle. Mol Biol Cell 18 3800 3809

49. BenantiJAMatyskielaMEMorganDOToczyskiDP 2009 Functionally distinct isoforms of Cik1 are differentially regulated by APC/C-mediated proteolysis. Mol Cell 33 581 590

50. JonesMHBachantJBCastilloARGiddingsTHJrWineyM 1999 Yeast Dam1p is required to maintain spindle integrity during mitosis and interacts with the Mps1p kinase. Mol Biol Cell 10 2377 2391

51. RenshawMJWardJJKanemakiMNatsumeKNedelecFJ 2010 Condensins promote chromosome recoiling during early anaphase to complete sister chromatid separation. Dev Cell 19 232 244

52. SharpDJRogersGCScholeyJM 2000 Cytoplasmic dynein is required for poleward chromosome movement during mitosis in Drosophila embryos. Nat Cell Biol 2 922 930

53. BaderJRVaughanKT 2010 Dynein at the kinetochore: Timing, Interactions and Functions. Semin Cell Dev Biol 21 269 275

54. MayerTUKapoorTMHaggartySJKingRWSchreiberSL 1999 Small molecule inhibitor of mitotic spindle bipolarity identified in a phenotype-based screen. Science 286 971 974

55. KapoorTMMayerTUCoughlinMLMitchisonTJ 2000 Probing spindle assembly mechanisms with monastrol, a small molecule inhibitor of the mitotic kinesin, Eg5. J Cell Biol 150 975 988

56. FamulskiJKChanGK 2007 Aurora B kinase-dependent recruitment of hZW10 and hROD to tensionless kinetochores. Curr Biol 17 2143 2149

57. LongtineMSMcKenzieA3rdDemariniDJShahNGWachA 1998 Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast 14 953 961

58. LiuHWangY 2006 The function and regulation of budding yeast Swe1 in response to interrupted DNA synthesis. Mol Biol Cell 17 2746 2756

59. LiuHLiangFJinFWangY 2008 The coordination of centromere replication, spindle formation, and kinetochore-microtubule interaction in budding yeast. PLoS Genet 4 e1000262 doi:10.1371/journal.pgen.1000262