Journal of Microbiology

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Characterization of a Putative F-box Motif in Ibd1p/Bfalp, a Spindle Checkpoint Regulator of Budding Yeast Saccharomyces cerevisiae

  • Lee, Kyum-Jung (Department of Bilochemistry and Institute of Life Science and Biotechnology, College of Science, Yonsei University) ;
  • Hyung-Seo (Department of Bilochemistry and Institute of Life Science and Biotechnology, College of Science, Yonsei University) ;
  • Kiwon Song (Department of Bilochemistry and Institute of Life Science and Biotechnology, College of Science, Yonsei University)
  • Published : 2001.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

During mitosis. the proper segregation of duplicated chromosomes is corrdinated by a spindle check-point. The bifurcated spindle checkpoint blocks cell cycle progression at metaphase by monitoring unattached kinetochores and inhibits mitotic exit in response to the misorientation of the mitotic spin- dle Ibd1p/Bfa1p is a spindle checkpoint regulator of budding yeast in the Bub2p checkpoint pathway for mitotic exit and its disruption abolishes mitotic arrest when proper organization of the mitotic spin-dls inhibited. Ibd1p/Bfa1p localizes to the spindle pole body, a microtublue-organizing center in yeast, and its overexpression arrests the cell cycle in 80% of cells with an enlarged budy at mitosis and in 20 % of cells with multiple buds. In this study, we found that the C-terminus of Ibd1p/Bfa1p phys-ically interacts with Skp1p, a key component of SCF (Skp1/cullin/F-box) complex for ubiquition-medi-ated proteolysis of cel cycle regulatores as well as an evolutionally conserved kinetochore protein for cell cycle progression. A putative F-box motif was found in the C-terminus of Ibd1p/Bfa1p and its function was investigated by making mutants of conserved residues in the motif. These Ibd1p/Bfa1p mutants of a putative F-box interacted with SKp1p in vitro by two-hybrid assays as wild type Ibd1p/Bfa1p. Also these Ibd1p/Bfa1p utants displayed the overexpression phenotypes of wild type Ibd1p, when over-expressed under inducible promoters . These results suggest that a putative F-box motif of Ibd1p/Bfa1p is not essential for the interaction with SKp1p and its function in mitotic exit and cytokinesis.

Keywords

References

  1. EMBO J. v.18 Sister chromatid separation and chromosome re-duplication are regulated by different mechanisms in response to spindle damage Alexandru G.;W. Zachariae;A. Schleiffer;K. Nasmyth
  2. Cell v.86 SKPI connects cell cycle regulators to the ubiquitin protecolysis machinery through a novel motif, the F-box Bai C.;P. Sen;K. Hofmann;L. Ma;M. Goebl;J.W. Harper;S.J. Elledge
  3. Cell v.102 A mechanism for coupling exit from mitosis to partitioning of the nucleus Bardin A.J.;R. Visintin;A. Amon
  4. Nat. Cell Biol. v.2 Anaphase spindle position is monitored by the BUB2 checkpoint Bloecher A.;G.M. Venturi;K. Tatchell
  5. Curr. Opin. Genet. Dev. v.10 Complexity in the spindle checkpoint Burke D.J.
  6. J. Cell. Biol. v.143 Spindle checkpoint protein Xmad1 recruits Xmad2 to unattached kinetochores Chen R.H.;A. Shevchenko;M. Mann;A.W. Murray
  7. Science v.274 Association of spindle assembly checkpoint component XMAD2 with unattached kinetochores Chen R.H.;J.C. Waters;E.D. Salmon;A.W. Murray
  8. Cell v.86 Budding yeast SKPI encodes an evolutionarily conserved kinetochore protein required for cell cycle progression Connelly C.;P. Hieter
  9. Prog. Biophys. Mol. Biol. v.72 The F-box: a new motif for ubiquitin dependent proteolysis in cell cycle regulation and signal transduction Craig K.L.;M. Tyers
  10. Mol. Cell v.2 Direct binding of CDC20 protein family members activates the anaphase-promoting complex in mitosis and G1 Fang G.;H. Yu;M.W. Kirschner
  11. Cell v.91 A complex of Cdc4, Skp1, and Cdc53/Cullin catalyze ubiquitination of the phosphorylated CDK inhiitor Sic1 Feldman R.M.;C.C. Corell;K.B. Kaplan;R.J. Deshaies
  12. EMBO J. v.18 A Bub2p-dependent spindle checkpoint pathway regulates the Dbf2p kinase in budding yeast Fesquet D.;P.J. Fitzpatrick;A.L. Johnson;K.M. Kramer;J.H. Toyn;L.H. Johnston
  13. J. Cell Biol. v.145 Budding yeast Bub2 is localized at spindle pole bodies and activates the mitotic checkpoint via a different pathway from Mad2 Fraschini R.;E. Formenti;G. Lucchini;S. Piatti
  14. Genes Dev. v.13 Components of an SCF ubiquitin ligase localize to the centrosome and regulate the centrosome duplication cycle Feed E.;K.R. Lacey;P. Huie;S.A. Lyapina;R.J. Deshaies;T. Stearns;P.K. Jackson
  15. Curr. Biol. v.8 Byr4 and Cde16 form a two-component GTPase-activating protein for the Spg1 GTPase that controls septation in fission yeast Furge K.A.;K. Wong;J. Armstrong;M. Balasubramanian;C.F. Albright
  16. Genetics v.157 The spindle checkpoint of the yeast Saccharomyces cerevisiae requires kinetochore function and maps to the CBF3 domain Gardner R.D.;A. Poddar;C. Yellman;P.A. Tavormina;M.C. Monteagudo;D.J. Burke
  17. Cell v.75 Cdi1, a human G1 and S phase protein phosphatase that associates with Cdk2 Gyuris J.;E. Golemis;H. Chertkov;R. Brent
  18. Cell v.106 Molecular analysis of Kinetochore-microtubule arrachment in budding yeast He X.;D.R. Rines;C.W. Espelin;P.K. Sorger
  19. Cell v.66 S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function Hoyt M.A.;L. Totis;B.T. Roberts
  20. Science v.279 Budding yeast Cdc20: a target of the spindle checkpoint Hwang L.H.;L. Lau;D. Smith;C.A. Mistrot;K.G. Hardwick
  21. Mol. Cells v.8 Byt4, a dosage-dependent regularot of cytokinesis in S. pombe, interacts with a possible small GTPase pathwat including Spg1 and Cdc16 Jwa M.;K. Song
  22. Science v.284 Rbx1, A component of the YHL tumor suppressor complex and SCF ubiquitin liagase Kamura T.;D.M. Koepp;M.N. Conrad;D. Skowyra;R.J. Moreland;O. Iliopoulos;W.S.Lane;W.G. Kaelin Jr,;S.J. Elledge;R.C. Conaway;J.W. Harper;J.W. Conaway
  23. Cell v.91 Regulating the yeast linetochore by ubiquitin-dependent degradation and Skp1p-mediated phosphorylation Kaplan K.B.;A.A. Hyman;P.K. Sorger
  24. Genome Biology v.1 The F-box protein family Kipreos E.;M. Pagano
  25. Biochimi. Biophysi. Acta v.1449 Ibd1p, a possible spindle pole body associated protein, regulates nuclear division and bud separation in Saccharomyces cerevisiae Lee J.;H. Hwang;J. Kim;K. Song
  26. Proc. Natl. Acad. Sci. v.96 Bifurcation of the mitotic checkpoint pathway in budding yeast Li R.
  27. Cell v.66 Feedback control of mitosis in budding yeast Li R.;A.W. Murray
  28. Mol. Cell Biol. v.16 Cdc53p acts in concert with Cdc4p and Cdc34p to control the G1-to-S-phase transition and identifies a conserved family of proteins Mathias N.;S.L. Johnson;M. Winey;A.E. Adams;L. Goetsch;J.R. Pringle;B. Byers;M.G. Goebl
  29. Proc. Natl. Acad. Sci. v.96 Retention of the BUB3 checkpoint protein on lagging chromosomes Martinez-Exposito M.J.;K.B. Kaplan;J. Copeland;P.K. Sorger
  30. Mol. Cell v.3 ROCI, a homolog of APC11, represents a family of cullin partners with an associated ubiquitin ligase activity Ohta T.;J.J. Michel;A.J. Schottelius;Y. Xiong
  31. Genes Dev. v.12 Cdc53 is a scaffold protein for multiple Cdc34/Sop1/F-box protein complexes that regulate cell division and methionine biosynthesis in yeast Patton E.E.;A.R. willems;D. Sa;L. Kuras;D. Thomas;K.L. Craig;M. Tyers
  32. Mol. Cell v.6 The Bub2p spindle checkpoint links nuclear migration with mitoic exit Pereira G.;T. Hofken;J. Grindlay;C. Manson;E. Schiebel
  33. Science v.284 Reconstitution of G1 cyclin ubiquitnation with complexes containing SCF Grrl and Rbx1 Skowyra D.;D.M. Koepp;T. Kamura;M.N. Conard;R.C. Conaway;J.W. Conaway;S.J. Elledge;J.W. Harper
  34. EMBO J. v.15 The Saccharomyces cerevisiae, as identified by in vitro reconstitution Stemmann O.;J. Lechner
  35. J. Cell Biol. v.142 The human homologue of Bub3 is required for kinetochore localization of Bub1 and a Mad3/Bub1-related protein kinase Taylor S.S.;E. Ha;F. McKeon
  36. Cell v.89 Kinetochore localization of murine Bub1 is required for normal mitotic and checkpoint response to spindle damage Taylor S.S.;F. McKeon
  37. J. Microbiol. v.39 Spindle checkpoint control in budding yeast Toh-e A.;Asakawa K.;S. Yoshida
  38. Cell v.86 Cdc53 targets phophorlated G1 cyclins for degradation by the ubiquitin proteolytic pathway Williams A.R.;S. Lanker;E.E. Patton;K.L. Crig;T.F. Nason;N. Mathias;R. Kobayashi;C. Wittenberg, and M. Tyers
  39. Science v.285 Identification of a vertebrate sister chromatid separation inhibitor involved in transformation and tumorigenesis Zou H.;T.J. McGarry;T. Bernal;M.A. Kirschner