Identification of host-derived subunits of phage SP RNA-dependent RNA polymerase (SP replicase).
Phage SP RNA-dependent RNA polymerase (SP replicase) was purified from Escherichia coli infected with RNA phage SP. The enzyme was found to be composed of four non-identical polypeptides, i.e. subunits I, II, III, and IV and molecular weights of 74,000, 69,000, 47,000, and 36,000 daltons, respectively. As in the case of phage Qbeta replicase, the largest polypeptide is identical with the ribosomal protein S1, and subunits III and IV with polypeptide chain elongation factors EF-Tu and EF-ts, respectively.. This is based on the behaviour of the subunits on SDS-polyacrylamide gel electrophoresis, isoelectric focusing and immunological cross-reaction. Subunits I, III, and IV of SP replicase are derived from the host cell, while subunit II is coded by phage RNA genome. The striking coincidence of the composition and entity of the structural components of SP replicase with those of Qbeta replicase may indicate the structural and functional requirements of host-derived polypeptides in RNA replicase. The binding activity of S1 (in 70S ribosome comples) to poly (U) is retained in SP replicase complex. In contrast, the GDP binding activity of EF-Tu is masked in SP replicase. It is concluded that S1 is required functionally whereas EF-Tu.EF-Ts are required structurally in RNA replicase.
D001426:Bacterial Proteins / Q000378:metabolism; D003090:Coliphages / Q000201:enzymology*; D004926:Escherichia coli / Q000201:enzymology*; D046911:Macromolecular Substances; D008970:Molecular Weight; D010445:Peptide Elongation Factors / Q000378:metabolism; D011072:Poly U / Q000378:metabolism; D011485:Protein Binding; D012316:RNA Nucleotidyltransferases / Q000302:isolation & purification*; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D012269:Ribosomal Proteins / Q000378:metabolism; D014764:Viral Proteins / Q000378:metabolism
D001426:Bacterial Proteins; D046911:Macromolecular Substances; D010445:Peptide Elongation Factors; D012269:Ribosomal Proteins; D014764:Viral Proteins; D011072:Poly U; D012316:RNA Nucleotidyltransferases; D012324:RNA-Dependent RNA Polymerase