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Sub1 and RPA associate with RNA polymerase II at different stages of transcription.

Single-stranded DNA-binding proteins play many roles in nucleic acid metabolism, but their importance during transcription remains unclear. Quantitative proteomic analysis of RNA polymerase II (RNApII) preinitiation complexes (PICs) identified Sub1 and the replication protein A complex (RPA), both of which bind single-stranded DNA (ssDNA). Sub1, homolog of mammalian coactivator PC4, exhibits strong genetic interactions with factors necessary for promoter melting. Sub1 localizes near the transcription bubble in vitro and binds to promoters in vivo dependent upon PIC assembly. In contrast, RPA localizes to transcribed regions of active genes, strongly correlated with transcribing RNApII but independently of replication. RFA1 interacts genetically with transcription elongation factor genes. Interestingly, RPA levels increase at active promoters in cells carrying a Sub1 deletion or ssDNA-binding mutant, suggesting competition for a common binding site. We propose that Sub1 and RPA interact with the nontemplate strand of RNApII complexes during initiation and elongation, respectively.

Pubmed ID: 22055186

Authors

  • Sikorski TW
  • Ficarro SB
  • Holik J
  • Kim T
  • Rando OJ
  • Marto JA
  • Buratowski S

Journal

Molecular cell

Publication Data

November 4, 2011

Associated Grants

  • Agency: NIGMS NIH HHS, Id: GM079205
  • Agency: NIGMS NIH HHS, Id: GM46498
  • Agency: NIGMS NIH HHS, Id: R01 GM046498
  • Agency: NIGMS NIH HHS, Id: R01 GM046498-19S1
  • Agency: NIGMS NIH HHS, Id: R01 GM046498-20
  • Agency: NIGMS NIH HHS, Id: R01 GM046498-21
  • Agency: NIGMS NIH HHS, Id: R01 GM056663
  • Agency: NIGMS NIH HHS, Id: R01 GM079205

Mesh Terms

  • Base Sequence
  • Binding Sites
  • DNA, Single-Stranded
  • DNA-Binding Proteins
  • Gene Expression Regulation, Fungal
  • Molecular Sequence Data
  • Peptide Chain Elongation, Translational
  • Peptide Chain Initiation, Translational
  • Promoter Regions, Genetic
  • Proteomics
  • RNA Polymerase II
  • Replication Protein A
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Transcription Initiation Site
  • Transcription, Genetic