Cloning of an RNA polymerase subunit cDNA of the fission yeast Schizosaccharomyces pombe by heterospecific complementation in Saccharomyces cerevisiae

G. V. Shpakovski, E. N. Lebedenko, P. Thuriaux

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

The rpb10 cDNA of the fission yeast Schizosaccharomyces pombe, encoding one of the five small subunits common to all three nuclear DNA-dependent RNA polymerases, was isolated from an expression cDNA library by two independent approaches: PCR-based screening and direct suppression by means of heterospecific complementation of a temperature-sensitive mutant defective in the corresponding gene of Saccharomyces cerevisiae. The cloned Sz. pombe cDNA encodes a protein Rpb10 of 71 amino acids with an M of 8275 Da, sharing 51 amino acids (71% identity) with the subunit ABC10β of RNA polymerases I-III from S. cerevisiae. All eukaryotic members of this protein family have the same general organization featuring two highly conserved motifs (RCFT/SCGK and RYCCRRM) around an atypical zinc finger and an additional invariant HVDLIEK motif toward the C-terminal end. The last motif is only characteristic for homologs from eukaryotes. In keeping with this remarkable structural conservation, the Sz. pombe cDNA also fully complemented a S. cerevisiae deletion mutant lacking subunit ABC10β (null allele rpb10-Δl::HlS3).

Original languageEnglish
Pages (from-to)100-106
Number of pages7
JournalRussian Journal of Bioorganic Chemistry
Volume23
Issue number2
Publication statusPublished - Feb 1997
Externally publishedYes

Keywords

  • Common subunits
  • Eukaryotic transcription
  • Fission yeast
  • Heterospecific complementation
  • RNA polymerases I-III
  • rpb10 gene
  • Suppressors
  • Zinc finger

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry

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