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Molecular and General Genetics MGG

, Volume 262, Issue 4–5, pp 749–757 | Cite as

Isolation and characterization of the fission yeast gene rpa42+, which encodes a subunit shared by RNA polymerases I and III

  • Y. Imazawa
  • K. Imai
  • A. Fukushima
  • K. Hisatake
  • M. Muramatsu
  • Y. Nogi
ORIGINAL PAPER

Abstract

Eukaryotic RNA polymerases I and III share two distinct α-related subunits that show limited homology to the α subunit of Escherichia coli RNA polymerase, which forms a homodimer to nucleate the assembly of prokaryotic RNA polymerase. To gain insight into the functions of α-related subunits in eukaryotes, we have previously identified the α-related small subunit RPA17 of RNA polymerase I (and III) in Schizosaccharomyces pombe, and have shown that it is a functional homolog of Saccharomyces cerevisiae AC19. In an extension of that study, we have now isolated and characterized rpa42+, which encodes the α-related large subunit RPA42 of S. pombe RNA polymerase I, by virtue of the fact that its product interacts with RPA17 in the yeast two-hybrid system. We have found that rpa42+ encodes a polypeptide with an apparent molecular mass of 42 kDa, which shows 58% identity to the AC40 subunit shared by RNA polymerases I and III in S. cerevisiae. Furthermore, we have shown that rpa42+ complements a temperature-sensitive mutation in RPC40 the gene that encodes AC40 in S. cerevisiae and which is essential for cell growth. Finally, we have shown that neither RPA42 nor RPA17 can self-associate. These results provide evidence that the two distinct α-related subunits, RPA42 and RPA17, of RNA polymerases I and III are functionally conserved between S. pombe and S. cerevisiae, and suggest that heterodimer formation between them is essential for the assembly of RNA polymerases I and III in eukaryotes.

Key words Fission yeast RNA polymerase I Two-hybrid system α-Subunit 

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Copyright information

© Springer-Verlag Berlin Heidelberg 1999

Authors and Affiliations

  • Y. Imazawa
    • 1
  • K. Imai
    • 1
  • A. Fukushima
    • 1
  • K. Hisatake
    • 1
  • M. Muramatsu
    • 1
  • Y. Nogi
    • 1
  1. 1.Department of Biochemistry, Saitama Medical School, 38 Morohongo, Moroyama, Iruma-Gun, Saitama 350-0095, Japan E-mail: yasunogi@saitama-med.ac.jp Fax: +81-492-949751JP

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