The amino-terminal domain of ELL transcription elongation factor is essential for ELL function in Schizosaccharomyces pombe

Microbiology (Reading). 2017 Nov;163(11):1641-1653. doi: 10.1099/mic.0.000554. Epub 2017 Oct 18.

Abstract

Transcriptional elongation is a critical step for regulating expression of protein-coding genes. Multiple transcription elongation factors have been identified in vitro, but the physiological roles of many of them are still not clearly understood. The ELL (Eleven nineteen Lysine rich Leukemia) family of transcription elongation factors are conserved from fission yeast to humans. Schizosaccharomyces pombe contains a single ELL homolog (SpELL) that is not essential for its survival. Therefore to gain insights into the in vivo cellular functions of SpELL, we identified phenotypes associated with deletion of ell1 in S. pombe. Our results demonstrate that SpELL is required for normal growth of S. pombe cells. Furthermore, cells lacking ell1+ exhibit a decrease in survival when exposed to DNA-damaging conditions, but their growth is not affected under environmental stress conditions. ELL orthologs in different organisms contain three conserved domains, an amino-terminal domain, a middle domain and a carboxyl-terminal domain. We also carried out an in vivo functional mapping of these conserved domains within S. pombe ELL and uncovered a critical role for its amino-terminus in regulating all its cellular functions, including growth under different conditions, transcriptional elongation potential and interaction with S. pombe EAF. Taken together our results suggest that the domain organization of ELL proteins is conserved across species, but the in vivo functions as well as the relationship between the various domains and roles of ELL show species-specific differences.

Keywords: EAF; ELL; RNA polymerase II; S. pombe; gene regulation; transcriptional elongation.

MeSH terms

  • Amino Acid Motifs / genetics
  • Amino Acid Sequence / genetics
  • DNA Damage / physiology
  • Gene Deletion
  • Protein Binding
  • Protein Domains / genetics
  • RNA Polymerase II / genetics
  • Schizosaccharomyces / genetics*
  • Schizosaccharomyces / growth & development
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces pombe Proteins / chemistry
  • Schizosaccharomyces pombe Proteins / genetics*
  • Schizosaccharomyces pombe Proteins / metabolism*
  • Sequence Homology, Amino Acid
  • Stress, Physiological
  • Structure-Activity Relationship
  • Transcription Factors / metabolism*
  • Transcription, Genetic / genetics*
  • Transcriptional Elongation Factors / chemistry
  • Transcriptional Elongation Factors / genetics*
  • Transcriptional Elongation Factors / metabolism*
  • Two-Hybrid System Techniques

Substances

  • Schizosaccharomyces pombe Proteins
  • Transcription Factors
  • Transcriptional Elongation Factors
  • eaf1 protein, S pombe
  • ell1 protein, S pombe
  • RNA Polymerase II