Spt5 Plays Vital Roles in the Control of Sense and Antisense Transcription Elongation

Mol Cell. 2017 Apr 6;66(1):77-88.e5. doi: 10.1016/j.molcel.2017.02.023. Epub 2017 Mar 30.

Abstract

Spt5 is an essential and conserved factor that functions in transcription and co-transcriptional processes. However, many aspects of the requirement for Spt5 in transcription are poorly understood. We have analyzed the consequences of Spt5 depletion in Schizosaccharomyces pombe using four genome-wide approaches. Our results demonstrate that Spt5 is crucial for a normal rate of RNA synthesis and distribution of RNAPII over transcription units. In the absence of Spt5, RNAPII localization changes dramatically, with reduced levels and a relative accumulation over the first ∼500 bp, suggesting that Spt5 is required for transcription past a barrier. Spt5 depletion also results in widespread antisense transcription initiating within this barrier region. Deletions of this region alter the distribution of RNAPII on the sense strand, suggesting that the barrier observed after Spt5 depletion is normally a site at which Spt5 stimulates elongation. Our results reveal a global requirement for Spt5 in transcription elongation.

Keywords: Spt5; antisense transcription; convergent antisense transcription; transcription elongation.

MeSH terms

  • Chromosomal Proteins, Non-Histone / genetics
  • Chromosomal Proteins, Non-Histone / metabolism*
  • Computational Biology
  • Databases, Genetic
  • Gene Expression Regulation, Fungal
  • Genome, Fungal
  • Genotype
  • Mutation
  • Phenotype
  • Promoter Regions, Genetic
  • RNA Polymerase II / metabolism
  • RNA Splicing
  • RNA, Antisense / biosynthesis*
  • RNA, Antisense / genetics
  • RNA, Fungal / biosynthesis*
  • RNA, Fungal / genetics
  • RNA, Messenger / biosynthesis*
  • RNA, Messenger / genetics
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism*
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism*
  • Schizosaccharomyces pombe Proteins / genetics
  • Schizosaccharomyces pombe Proteins / metabolism*
  • Time Factors
  • Transcription Elongation, Genetic*
  • Transcriptional Elongation Factors / genetics
  • Transcriptional Elongation Factors / metabolism*

Substances

  • Chromosomal Proteins, Non-Histone
  • RNA, Antisense
  • RNA, Fungal
  • RNA, Messenger
  • Schizosaccharomyces pombe Proteins
  • Spt5 protein, S pombe
  • Transcriptional Elongation Factors
  • SPT5 transcriptional elongation factor
  • RNA Polymerase II