The N-terminal amino acid sequence is essential for foot-and-mouth disease virus replicase activity

Braz J Med Biol Res. 1992;25(7):659-66.

Abstract

1. Foot-and-mouth disease virus replicase was expressed by fusing its cDNA to the OmpA signal peptide coding sequence present in the pIN-III ompA series vectors. 2. Two constructions were developed to express either a full-length or truncated enzyme lacking the 20 amino acids at the N-terminal end. Bacterial extracts expressing the recombinant proteins were submitted to SDS-PAGE and the presence of the replicase was revealed by immunoblotting. The truncated form exhibited a higher mobility and the relative positions of the proteins show that the signal peptide was removed. 3. The biological activity of these two molecules was tested using a poly(A)-dependent oligo(U)-primed poly(U)-polymerase assay. The full-length replicase is active. The aminoterminal truncated enzyme had 0.02% activity of the intact one. 4. This result indicates the importance of the twenty N-terminal amino acids for the activity of FMDV RNA-dependent RNA polymerase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aphthovirus / enzymology*
  • Aphthovirus / physiology
  • Base Sequence
  • DNA Replication*
  • DNA, Viral / metabolism*
  • DNA-Directed DNA Polymerase / analysis
  • DNA-Directed DNA Polymerase / metabolism*
  • Escherichia coli / genetics
  • Molecular Sequence Data
  • Plasmids
  • Structure-Activity Relationship
  • Virus Replication*

Substances

  • DNA, Viral
  • DNA replicase
  • DNA-Directed DNA Polymerase