Saccharomyces cerevisiae centromere RNA is negatively regulated by Cbf1 and its unscheduled synthesis impacts CenH3 binding

Chi Fu Chen, Thomas J. Pohl, Angela Chan, Joshua S. Slocum, Virginia A. Zakian

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Two common features of centromeres are their transcription into noncoding centromere RNAs (cen-RNAs) and their assembly into nucleosomes that contain a centromere-specific histone H3 (cenH3). Here, we show that Saccharomyces cerevisiae cen- RNA was present in low amounts in wild-type (WT) cells, and that its appearance was tightly cell cycle-regulated, appearing and disappearing in a narrow window in S phase after centromere replication. In cells lacking Cbf1, a centromere-binding protein, cen-RNA was 5-12 times more abundant throughout the cell cycle. In WT cells, cen-RNA appearance occurred at the same time as loss of Cbf1's centromere binding, arguing that the physical presence of Cbf1 inhibits cen-RNA production. Binding of the Pif1 DNA helicase, which happens in mid-late S phase, occurred at about the same time as Cbf1 loss from the centromere, suggesting that Pif1 may facilitate this loss by its known ability to displace proteins from DNA. Cen-RNAs were more abundant in rnh1D cells but only in mid-late S phase. However, fork pausing at centromeres was not elevated in rnh1D cells but rather was due to centromere-binding proteins, including Cbf1. Strains with increased cen-RNA lost centromere plasmids at elevated rates. In cbf1D cells, where both the levels and the cell cycle-regulated appearance of cen-RNA were disrupted, the timing and levels of cenH3 centromere binding were perturbed. Thus, cen-RNAs are highly regulated, and disruption of this regulation correlates with changes in centromere structure and function.

Original languageEnglish (US)
Pages (from-to)465-479
Number of pages15
JournalGenetics
Volume213
Issue number2
DOIs
StatePublished - 2019

All Science Journal Classification (ASJC) codes

  • Genetics

Keywords

  • Cbf1
  • Centromere
  • Centromere RNA
  • Cse4

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