The role of the carboxyterminal domain of RNA polymerase II in regulating origins of DNA replication in Saccharomyces cerevisiae

Genetics. 2002 Nov;162(3):1117-29. doi: 10.1093/genetics/162.3.1117.

Abstract

MCM (minichromosome maintenance) proteins function as a replication licensing factor (RLF-M), which contributes to limiting initiation of DNA replication to once per cell cycle. In the present study we show that a truncation of the pol II CTD in a S. cerevisiae strain harboring a mutation in mcm5 partially reverses its ts phenotype and improves maintenance of CEN/ARS minichromosomes. We correlate this phenotype to effects on DNA replication rather than to effects on transcription or specific gene expression. We also demonstrate that a similar truncation of the CTD reduces minichromosome stability and impairs stimulation of DNA replication by trans-activators and that tethering of recombinant pol II CTD to an origin of replication has a significant stimulatory effect on minichromosome stability. Furthermore, we show that pol II is recruited to ARS1. We propose that in S. cerevisiae a mechanism of coordinating pol II transcription and DNA replication is mediated by the CTD of pol II.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Cycle Proteins / genetics
  • Cell Cycle Proteins / metabolism
  • DNA / metabolism
  • DNA Replication
  • DNA-Binding Proteins / metabolism
  • Gene Expression Regulation, Fungal / physiology*
  • Mutation
  • Protein Structure, Tertiary
  • RNA Polymerase II / genetics
  • RNA Polymerase II / metabolism*
  • Replication Origin*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / metabolism
  • Schizosaccharomyces pombe Proteins
  • Transcription Factors / metabolism
  • Transcription, Genetic

Substances

  • ABF1 protein, S cerevisiae
  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • GAL4 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Schizosaccharomyces pombe Proteins
  • Transcription Factors
  • mcm5 protein, S pombe
  • DNA
  • RNA Polymerase II