Quantitative Proteomic Analysis of Purified Yeast Kinetochores Identifies a PP1 Regulatory Subunit

Overview

Journal Genes Dev

Specialty Molecular Biology

Date 2009 Dec 2

PMID 19948764

Citations 76

Authors

Bungo Akiyoshi

Christian R Nelson

Jeffrey A Ranish

Sue Biggins

Affiliations

Soon will be listed here.

Abstract

The kinetochore is a macromolecular complex that controls chromosome segregation and cell cycle progression. When sister kinetochores make bioriented attachments to microtubules from opposite poles, the spindle checkpoint is silenced. Biorientation and the spindle checkpoint are regulated by a balance between the Ipl1/Aurora B protein kinase and the opposing activity of protein phosphatase I (PP1). However, little is known about the regulation of PP1 localization and activity at the kinetochore. Here, we developed a method to purify centromere-bound kinetochores and used quantitative proteomics to identify the Fin1 protein as a PP1 regulatory subunit. The Fin1/PP1 complex is regulated by phosphorylation and 14-3-3 protein binding. When Fin1 is mislocalized, bipolar spindles fail to assemble but the spindle checkpoint is inappropriately silenced due to PP1 activity. These data suggest that Fin1 is a PP1 regulatory subunit whose spatial and temporal activity must be precisely controlled to ensure genomic stability.

Citing Articles

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References

Sandall S, Severin F, McLeod I, Yates 3rd J, Oegema K, Hyman A . A Bir1-Sli15 complex connects centromeres to microtubules and is required to sense kinetochore tension. Cell. 2006; 127(6):1179-91. PMC: 2265205. DOI: 10.1016/j.cell.2006.09.049. View

Wu J, Guo J, Tang W, Yang C, Freel C, Chen C . PP1-mediated dephosphorylation of phosphoproteins at mitotic exit is controlled by inhibitor-1 and PP1 phosphorylation. Nat Cell Biol. 2009; 11(5):644-51. PMC: 2788612. DOI: 10.1038/ncb1871. View

Lin S, Riggs A . The general affinity of lac repressor for E. coli DNA: implications for gene regulation in procaryotes and eucaryotes. Cell. 1975; 4(2):107-11. DOI: 10.1016/0092-8674(75)90116-6. View

Pinsky B, Kotwaliwale C, Tatsutani S, Breed C, Biggins S . Glc7/protein phosphatase 1 regulatory subunits can oppose the Ipl1/aurora protein kinase by redistributing Glc7. Mol Cell Biol. 2006; 26(7):2648-60. PMC: 1430313. DOI: 10.1128/MCB.26.7.2648-2660.2006. View

Buvelot S, Tatsutani S, Vermaak D, Biggins S . The budding yeast Ipl1/Aurora protein kinase regulates mitotic spindle disassembly. J Cell Biol. 2003; 160(3):329-39. PMC: 2172676. DOI: 10.1083/jcb.200209018. View

Pinsky B, Kung C, Shokat K, Biggins S . The Ipl1-Aurora protein kinase activates the spindle checkpoint by creating unattached kinetochores. Nat Cell Biol. 2005; 8(1):78-83. DOI: 10.1038/ncb1341. View

De Wulf P, McAinsh A, Sorger P . Hierarchical assembly of the budding yeast kinetochore from multiple subcomplexes. Genes Dev. 2003; 17(23):2902-21. PMC: 289150. DOI: 10.1101/gad.1144403. View

Biggins S, Murray A . The budding yeast protein kinase Ipl1/Aurora allows the absence of tension to activate the spindle checkpoint. Genes Dev. 2001; 15(23):3118-29. PMC: 312839. DOI: 10.1101/gad.934801. View

Cohen P . Protein phosphatase 1--targeted in many directions. J Cell Sci. 2002; 115(Pt 2):241-56. DOI: 10.1242/jcs.115.2.241. View

10.

Clarke L, Carbon J . Isolation of a yeast centromere and construction of functional small circular chromosomes. Nature. 1980; 287(5782):504-9. DOI: 10.1038/287504a0. View

11.

Emanuele M, Lan W, Jwa M, Miller S, Chan C, Stukenberg P . Aurora B kinase and protein phosphatase 1 have opposing roles in modulating kinetochore assembly. J Cell Biol. 2008; 181(2):241-54. PMC: 2315672. DOI: 10.1083/jcb.200710019. View

12.

Bloecher A, Tatchell K . Dynamic localization of protein phosphatase type 1 in the mitotic cell cycle of Saccharomyces cerevisiae. J Cell Biol. 2000; 149(1):125-40. PMC: 2175104. DOI: 10.1083/jcb.149.1.125. View

13.

Surana U, Amon A, Dowzer C, McGrew J, Byers B, Nasmyth K . Destruction of the CDC28/CLB mitotic kinase is not required for the metaphase to anaphase transition in budding yeast. EMBO J. 1993; 12(5):1969-78. PMC: 413418. DOI: 10.1002/j.1460-2075.1993.tb05846.x. View

14.

Furuyama S, Biggins S . Centromere identity is specified by a single centromeric nucleosome in budding yeast. Proc Natl Acad Sci U S A. 2007; 104(37):14706-11. PMC: 1976213. DOI: 10.1073/pnas.0706985104. View

15.

Sassoon I, Severin F, Andrews P, Taba M, Kaplan K, Ashford A . Regulation of Saccharomyces cerevisiae kinetochores by the type 1 phosphatase Glc7p. Genes Dev. 1999; 13(5):545-55. PMC: 316511. DOI: 10.1101/gad.13.5.545. View

16.

Ortiz J, Stemmann O, Rank S, Lechner J . A putative protein complex consisting of Ctf19, Mcm21, and Okp1 represents a missing link in the budding yeast kinetochore. Genes Dev. 1999; 13(9):1140-55. PMC: 316948. DOI: 10.1101/gad.13.9.1140. View

17.

Hendrickx A, Beullens M, Ceulemans H, Abt T, Van Eynde A, Nicolaescu E . Docking motif-guided mapping of the interactome of protein phosphatase-1. Chem Biol. 2009; 16(4):365-71. DOI: 10.1016/j.chembiol.2009.02.012. View

18.

Mayordomo I, Sanz P . The Saccharomyces cerevisiae 14-3-3 protein Bmh2 is required for regulation of the phosphorylation status of Fin1, a novel intermediate filament protein. Biochem J. 2002; 365(Pt 1):51-6. PMC: 1222645. DOI: 10.1042/BJ20020053. View

19.

Pinsky B, Tatsutani S, Collins K, Biggins S . An Mtw1 complex promotes kinetochore biorientation that is monitored by the Ipl1/Aurora protein kinase. Dev Cell. 2003; 5(5):735-45. DOI: 10.1016/s1534-5807(03)00322-8. View

20.

Kakiuchi K, Yamauchi Y, Taoka M, Iwago M, Fujita T, Ito T . Proteomic analysis of in vivo 14-3-3 interactions in the yeast Saccharomyces cerevisiae. Biochemistry. 2007; 46(26):7781-92. DOI: 10.1021/bi700501t. View