[Cloning of cDNA for RNA polymerase subunit from the fission yeast Schizosaccharomyces pombe by heterospecific complementation in Saccharomyces cerevisiae]

Bioorg Khim. 1997 Feb;23(2):110-7.
[Article in Russian]

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 8,275 Da, sharing 51 amino acids (71% identity) with the subunit ABC10 beta 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 characteristics 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 ABC10 beta (null allele rpb10-delta 1::HIS3).

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / genetics*
  • DNA-Directed RNA Polymerases / genetics*
  • Genes, Fungal
  • Genetic Complementation Test
  • Molecular Sequence Data
  • Plasmids
  • Polymerase Chain Reaction
  • Saccharomyces cerevisiae / genetics*
  • Schizosaccharomyces / enzymology*
  • Schizosaccharomyces / genetics

Substances

  • DNA, Complementary
  • DNA-Directed RNA Polymerases

Associated data

  • GENBANK/AF027818
  • GENBANK/U80219