Rtt107/Esc4 binds silent chromatin and DNA repair proteins using different BRCT motifs

David C. Zappulla, Arindel S Maharaj, Jessica J. Connelly, Rebecca A. Jockusch, Rolf Sternglanz

Research output: Contribution to journalArticle

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Abstract

Background: By screening a plasmid library for proteins that could cause silencing when targeted to the HMR locus in Saccharomyces cerevisiae, we previously reported the identification of Rtt107/Esc4 based on its ability to establish silent chromatin. In this study we aimed to determine the mechanism of Rtt107/Esc4 targeted silencing and also learn more about its biological functions. Results: Targeted silencing by Rtt107/Esc4 was dependent on the SIR genes, which encode obligatory structural and enzymatic components of yeast silent chromatin. Based on its sequence, Rtt107/Esc4 was predicted to contain six BRCT motifs. This motif, originally identified in the human breast tumor suppressor gene BRCAI, is a protein interaction domain. The targeted silencing activity of Rtt107/Esc4 resided within the C-terminal two BRCT motifs, and this region of the protein bound to Sir3 in two-hybrid tests. Deletion of RTT107/ESC4 caused sensitivity to the DNA damaging agent MMS as well as to hydroxyurea. A two-hybrid screen showed that the N-terminal BRCT motifs of Rtt107/Esc4 bound to Slx4, a protein previously shown to be involved in DNA repair and required for viability in a strain lacking the DNA helicase Sgs1. Like SLX genes, RTT107ESC4 interacted genetically with SGS1; esc4Δ sgs1Δ mutants were viable, but exhibited a slow-growth phenotype and also a synergistic DNA repair defect. Conclusion: Rtt107/Esc4 binds to the silencing protein Sir3 and the DNA repair protein Slx4 via different BRCT motifs, thus providing a bridge linking silent chromatin to DNA repair enzymes.

Original languageEnglish (US)
Article number40
JournalBMC Molecular Biology
Volume7
DOIs
StatePublished - Nov 9 2006
Externally publishedYes

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DNA Repair
Chromatin
Repair
DNA
Genes
Proteins
Yeast
Protein Interaction Domains and Motifs
DNA Repair Enzymes
DNA Helicases
Amino Acid Motifs
Hydroxyurea
Tumor Suppressor Genes
Saccharomyces cerevisiae
Plasmids
Yeasts
Breast Neoplasms
Tumors
Phenotype
Screening

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Rtt107/Esc4 binds silent chromatin and DNA repair proteins using different BRCT motifs. / Zappulla, David C.; Maharaj, Arindel S; Connelly, Jessica J.; Jockusch, Rebecca A.; Sternglanz, Rolf.

In: BMC Molecular Biology, Vol. 7, 40, 09.11.2006.

Research output: Contribution to journalArticle

Zappulla, David C. ; Maharaj, Arindel S ; Connelly, Jessica J. ; Jockusch, Rebecca A. ; Sternglanz, Rolf. / Rtt107/Esc4 binds silent chromatin and DNA repair proteins using different BRCT motifs. In: BMC Molecular Biology. 2006 ; Vol. 7.
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