A role for CENP-A/Cse4 phosphorylation on serine 33 in deposition at the centromere

Gesine Hoffmann, Anke Samel-Pommerencke, Jan Weber, Alessandro Cuomo, Tiziana Bonaldi, Ann E Ehrenhofer-Murray

Research output: Contribution to journalArticle

Abstract

Centromeres are the sites of assembly of the kinetochore, which connect the chromatids to the microtubules for sister chromatid segregation during cell division. Centromeres are characterized by the presence of the histone H3 variant CENP-A (termed Cse4 in Saccharomyces cerevisiae). Here, we investigated the function of serine 33 phosphorylation of Cse4 (Cse4-S33ph) in S. cerevisiae, which lies within the essential N-terminal domain (END) of the extended Cse4 N-terminus. Significantly, we identified histone H4-K5, 8, 12R to cause a temperature-sensitive growth defect with mutations in Cse4-S33 and sensitivity to nocodazole and hydroxyurea. Furthermore, the absence of Cse4-S33ph reduced the levels of Cse4 at centromeric sequences, suggesting that Cse4 deposition is defective in the absence of S33 phosphorylation. We furthermore identified synthetic genetic interactions with histone H2A-E57A and H2A-L66A, which both cause a reduced interaction with the histone chaperone FACT and reduced H2A/H2B levels in chromatin, again supporting the notion that a combined defect of H2A/H2B and Cse4 deposition causes centromeric defects. Altogether, our data highlight the importance of correct histone deposition in building a functional centromeric nucleosome and suggests a role for Cse4-S33ph in this process.

Original languageEnglish
JournalFEMS Yeast Research
Volume18
Issue number1
DOIs
Publication statusPublished - Feb 1 2018

Fingerprint

Centromere
Histones
Serine
Phosphorylation
Chromatids
Saccharomyces cerevisiae
Histone Chaperones
Nocodazole
Kinetochores
Hydroxyurea
Nucleosomes
Microtubules
Cell Division
Chromatin
Mutation
Temperature
centromere protein A
Growth

Keywords

  • Centromere/genetics
  • Centromere Protein A/chemistry
  • Chromosomal Proteins, Non-Histone/chemistry
  • DNA Methylation
  • DNA-Binding Proteins/chemistry
  • Mass Spectrometry
  • Mutation
  • Phosphorylation
  • Saccharomyces cerevisiae Proteins/chemistry
  • Serine/metabolism

Cite this

A role for CENP-A/Cse4 phosphorylation on serine 33 in deposition at the centromere. / Hoffmann, Gesine; Samel-Pommerencke, Anke; Weber, Jan; Cuomo, Alessandro; Bonaldi, Tiziana; Ehrenhofer-Murray, Ann E.

In: FEMS Yeast Research, Vol. 18, No. 1, 01.02.2018.

Research output: Contribution to journalArticle

Hoffmann, Gesine ; Samel-Pommerencke, Anke ; Weber, Jan ; Cuomo, Alessandro ; Bonaldi, Tiziana ; Ehrenhofer-Murray, Ann E. / A role for CENP-A/Cse4 phosphorylation on serine 33 in deposition at the centromere. In: FEMS Yeast Research. 2018 ; Vol. 18, No. 1.
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abstract = "Centromeres are the sites of assembly of the kinetochore, which connect the chromatids to the microtubules for sister chromatid segregation during cell division. Centromeres are characterized by the presence of the histone H3 variant CENP-A (termed Cse4 in Saccharomyces cerevisiae). Here, we investigated the function of serine 33 phosphorylation of Cse4 (Cse4-S33ph) in S. cerevisiae, which lies within the essential N-terminal domain (END) of the extended Cse4 N-terminus. Significantly, we identified histone H4-K5, 8, 12R to cause a temperature-sensitive growth defect with mutations in Cse4-S33 and sensitivity to nocodazole and hydroxyurea. Furthermore, the absence of Cse4-S33ph reduced the levels of Cse4 at centromeric sequences, suggesting that Cse4 deposition is defective in the absence of S33 phosphorylation. We furthermore identified synthetic genetic interactions with histone H2A-E57A and H2A-L66A, which both cause a reduced interaction with the histone chaperone FACT and reduced H2A/H2B levels in chromatin, again supporting the notion that a combined defect of H2A/H2B and Cse4 deposition causes centromeric defects. Altogether, our data highlight the importance of correct histone deposition in building a functional centromeric nucleosome and suggests a role for Cse4-S33ph in this process.",
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T1 - A role for CENP-A/Cse4 phosphorylation on serine 33 in deposition at the centromere

AU - Hoffmann, Gesine

AU - Samel-Pommerencke, Anke

AU - Weber, Jan

AU - Cuomo, Alessandro

AU - Bonaldi, Tiziana

AU - Ehrenhofer-Murray, Ann E

N1 - © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

PY - 2018/2/1

Y1 - 2018/2/1

N2 - Centromeres are the sites of assembly of the kinetochore, which connect the chromatids to the microtubules for sister chromatid segregation during cell division. Centromeres are characterized by the presence of the histone H3 variant CENP-A (termed Cse4 in Saccharomyces cerevisiae). Here, we investigated the function of serine 33 phosphorylation of Cse4 (Cse4-S33ph) in S. cerevisiae, which lies within the essential N-terminal domain (END) of the extended Cse4 N-terminus. Significantly, we identified histone H4-K5, 8, 12R to cause a temperature-sensitive growth defect with mutations in Cse4-S33 and sensitivity to nocodazole and hydroxyurea. Furthermore, the absence of Cse4-S33ph reduced the levels of Cse4 at centromeric sequences, suggesting that Cse4 deposition is defective in the absence of S33 phosphorylation. We furthermore identified synthetic genetic interactions with histone H2A-E57A and H2A-L66A, which both cause a reduced interaction with the histone chaperone FACT and reduced H2A/H2B levels in chromatin, again supporting the notion that a combined defect of H2A/H2B and Cse4 deposition causes centromeric defects. Altogether, our data highlight the importance of correct histone deposition in building a functional centromeric nucleosome and suggests a role for Cse4-S33ph in this process.

AB - Centromeres are the sites of assembly of the kinetochore, which connect the chromatids to the microtubules for sister chromatid segregation during cell division. Centromeres are characterized by the presence of the histone H3 variant CENP-A (termed Cse4 in Saccharomyces cerevisiae). Here, we investigated the function of serine 33 phosphorylation of Cse4 (Cse4-S33ph) in S. cerevisiae, which lies within the essential N-terminal domain (END) of the extended Cse4 N-terminus. Significantly, we identified histone H4-K5, 8, 12R to cause a temperature-sensitive growth defect with mutations in Cse4-S33 and sensitivity to nocodazole and hydroxyurea. Furthermore, the absence of Cse4-S33ph reduced the levels of Cse4 at centromeric sequences, suggesting that Cse4 deposition is defective in the absence of S33 phosphorylation. We furthermore identified synthetic genetic interactions with histone H2A-E57A and H2A-L66A, which both cause a reduced interaction with the histone chaperone FACT and reduced H2A/H2B levels in chromatin, again supporting the notion that a combined defect of H2A/H2B and Cse4 deposition causes centromeric defects. Altogether, our data highlight the importance of correct histone deposition in building a functional centromeric nucleosome and suggests a role for Cse4-S33ph in this process.

KW - Centromere/genetics

KW - Centromere Protein A/chemistry

KW - Chromosomal Proteins, Non-Histone/chemistry

KW - DNA Methylation

KW - DNA-Binding Proteins/chemistry

KW - Mass Spectrometry

KW - Mutation

KW - Phosphorylation

KW - Saccharomyces cerevisiae Proteins/chemistry

KW - Serine/metabolism

U2 - 10.1093/femsyr/fox094

DO - 10.1093/femsyr/fox094

M3 - Article

VL - 18

JO - FEMS Yeast Research

JF - FEMS Yeast Research

SN - 1567-1356

IS - 1

ER -