Phosphorylation by Casein Kinase 2 Facilitates Psh1 Protein-assisted Degradation of Cse4 Protein

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2014 Sep 4

PMID 25183013

Citations 20

Authors

Geetha S Hewawasam

Mark Mattingly

Swaminathan Venkatesh

Ying Zhang

Laurence Florens

Jerry L Workman

Jennifer L Gerton

Affiliations

Soon will be listed here.

Abstract

Cse4 is the centromeric histone H3 variant in budding yeast. Psh1 is an E3 ubiquitin ligase that controls Cse4 levels through proteolysis. Here we report that Psh1 is phosphorylated by the Cka2 subunit of casein kinase 2 (CK2) to promote its E3 activity for Cse4. Deletion of CKA2 significantly stabilized Cse4. Consistent with phosphorylation promoting the activity of Psh1, Cse4 was stabilized in a Psh1 phosphodepleted mutant strain in which the major phosphorylation sites were changed to alanines. Phosphorylation of Psh1 did not control Psh1-Cse4 or Psh1-Ubc3(E2) interactions. Although Cse4 was highly stabilized in a cka2Δ strain, mislocalization of Cse4 was mild, suggesting that Cse4 misincorporation was prevented by the intact Psh1-Cse4 association. Supporting this idea, Psh1 was also stabilized in a cka2Δ strain. Collectively our data suggest that phosphorylation is crucial in Psh1-assisted control of Cse4 levels and that the Psh1-Cse4 association itself functions to prevent Cse4 misincorporation.

Citing Articles

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Interaction of histone H4 with Cse4 facilitates conformational changes in Cse4 for its sumoylation and mislocalization.

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References

Pardo P, MURRAY P, Walz K, Franco L, PASSERON S . In vivo and in vitro phosphorylation of the alpha 7/PRS1 subunit of Saccharomyces cerevisiae 20 S proteasome: in vitro phosphorylation by protein kinase CK2 is absolutely dependent on polylysine. Arch Biochem Biophys. 1998; 349(2):397-401. DOI: 10.1006/abbi.1997.0466. View

Camahort R, Li B, Florens L, Swanson S, Washburn M, Gerton J . Scm3 is essential to recruit the histone h3 variant cse4 to centromeres and to maintain a functional kinetochore. Mol Cell. 2007; 26(6):853-65. DOI: 10.1016/j.molcel.2007.05.013. View

Willems A, Schwab M, Tyers M . A hitchhiker's guide to the cullin ubiquitin ligases: SCF and its kin. Biochim Biophys Acta. 2004; 1695(1-3):133-70. DOI: 10.1016/j.bbamcr.2004.09.027. View

Meggio F, Pinna L . One-thousand-and-one substrates of protein kinase CK2?. FASEB J. 2003; 17(3):349-68. DOI: 10.1096/fj.02-0473rev. View

Krogan N, Kim M, Ahn S, Zhong G, Kobor M, Cagney G . RNA polymerase II elongation factors of Saccharomyces cerevisiae: a targeted proteomics approach. Mol Cell Biol. 2002; 22(20):6979-92. PMC: 139818. DOI: 10.1128/MCB.22.20.6979-6992.2002. View

Tomonaga T, Matsushita K, Yamaguchi S, Oohashi T, Shimada H, Ochiai T . Overexpression and mistargeting of centromere protein-A in human primary colorectal cancer. Cancer Res. 2003; 63(13):3511-6. View

Van Hooser A, Ouspenski I, Gregson H, STARR D, Yen T, Goldberg M . Specification of kinetochore-forming chromatin by the histone H3 variant CENP-A. J Cell Sci. 2001; 114(Pt 19):3529-42. DOI: 10.1242/jcs.114.19.3529. View

Sadowski M, Mawson A, Baker R, Sarcevic B . Cdc34 C-terminal tail phosphorylation regulates Skp1/cullin/F-box (SCF)-mediated ubiquitination and cell cycle progression. Biochem J. 2007; 405(3):569-81. PMC: 2267305. DOI: 10.1042/BJ20061812. View

Collins K, Camahort R, Seidel C, Gerton J, Biggins S . The overexpression of a Saccharomyces cerevisiae centromeric histone H3 variant mutant protein leads to a defect in kinetochore biorientation. Genetics. 2006; 175(2):513-25. PMC: 1800591. DOI: 10.1534/genetics.106.064410. View

10.

Ludemann R, Lerea K, Etlinger J . Copurification of casein kinase II with 20 S proteasomes and phosphorylation of a 30-kDa proteasome subunit. J Biol Chem. 1993; 268(23):17413-7. View

11.

Au W, Crisp M, DeLuca S, Rando O, Basrai M . Altered dosage and mislocalization of histone H3 and Cse4p lead to chromosome loss in Saccharomyces cerevisiae. Genetics. 2008; 179(1):263-75. PMC: 2390605. DOI: 10.1534/genetics.108.088518. View

12.

Zybailov B, Mosley A, Sardiu M, Coleman M, Florens L, Washburn M . Statistical analysis of membrane proteome expression changes in Saccharomyces cerevisiae. J Proteome Res. 2006; 5(9):2339-47. DOI: 10.1021/pr060161n. View

13.

London N, Ceto S, Ranish J, Biggins S . Phosphoregulation of Spc105 by Mps1 and PP1 regulates Bub1 localization to kinetochores. Curr Biol. 2012; 22(10):900-6. PMC: 3723133. DOI: 10.1016/j.cub.2012.03.052. 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.

Hyman A, Sorger P . Structure and function of kinetochores in budding yeast. Annu Rev Cell Dev Biol. 1995; 11:471-95. DOI: 10.1146/annurev.cb.11.110195.002351. View

16.

Moreno-Moreno O, Medina-Giro S, Torras-Llort M, Azorin F . The F box protein partner of paired regulates stability of Drosophila centromeric histone H3, CenH3(CID). Curr Biol. 2011; 21(17):1488-93. DOI: 10.1016/j.cub.2011.07.041. View

17.

Belle A, Tanay A, Bitincka L, Shamir R, OShea E . Quantification of protein half-lives in the budding yeast proteome. Proc Natl Acad Sci U S A. 2006; 103(35):13004-9. PMC: 1550773. DOI: 10.1073/pnas.0605420103. View

18.

Puig O, Caspary F, Rigaut G, Rutz B, Bouveret E, Wilm M . The tandem affinity purification (TAP) method: a general procedure of protein complex purification. Methods. 2001; 24(3):218-29. DOI: 10.1006/meth.2001.1183. View

19.

Collins K, Furuyama S, Biggins S . Proteolysis contributes to the exclusive centromere localization of the yeast Cse4/CENP-A histone H3 variant. Curr Biol. 2004; 14(21):1968-72. DOI: 10.1016/j.cub.2004.10.024. View

20.

Hewawasam G, Shivaraju M, Mattingly M, Venkatesh S, Martin-Brown S, Florens L . Psh1 is an E3 ubiquitin ligase that targets the centromeric histone variant Cse4. Mol Cell. 2010; 40(3):444-54. PMC: 2998187. DOI: 10.1016/j.molcel.2010.10.014. View