Three tuf-like genes in the kirromycin producer Streptomyces ramocissimus

Microbiology (Reading). 1994 Apr;140 ( Pt 4):983-98. doi: 10.1099/00221287-140-4-983.

Abstract

We have identified, cloned and sequenced three tuf-like genes from Streptomyces ramocissimus (Sr.), the producer of the antibiotic kirromycin which inhibits protein synthesis by binding the polypeptide chain elongation factor EF-Tu. The tuf-1 gene encodes a protein with 71% amino acid residues identical to the well characterized elongation factor Tu of Escherichia coli (Ec.EF-Tu). The genetic location of tuf-1 downstream of a fus homologue and the in vitro activity of Sr.EF-Tu1 show that tuf-1 encodes a genuine EF-Tu. The putative Sr.EF-Tu2 and Sr.EF-Tu3 proteins are 69% and 63% identical to Ec.EF-Tu. Homologues of tuf-1 and tuf-3 were detected in all five Streptomyces strains investigated, but tuf-2 was found in S. ramocissimus only. The three tuf genes were expressed in E. coli and used to produce polyclonal antibodies. Western blot analysis showed that Sr.EF-Tu1 was present at all times under kirromycin production conditions in submerged and surface-grown cultures of S. ramocissimus and in germinating spores. The expression of tuf-2 and tuf-3 was, however, below the detection level. Surprisingly, Sr.EF-Tu1 was kirromycin sensitive, which excludes the possibility that EF-Tu is involved in the kirromycin resistance of S. ramocissimus.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Bacterial Proteins / biosynthesis
  • Bacterial Proteins / genetics*
  • Base Sequence
  • Cloning, Molecular
  • Drug Resistance, Microbial / genetics
  • Escherichia coli / genetics
  • Gene Expression Regulation
  • Genes, Bacterial*
  • Molecular Sequence Data
  • Peptide Elongation Factor Tu / biosynthesis
  • Peptide Elongation Factor Tu / genetics*
  • Pyridones / metabolism
  • Pyridones / pharmacology
  • Recombinant Fusion Proteins / biosynthesis
  • Sequence Alignment
  • Sequence Homology
  • Species Specificity
  • Spores, Bacterial
  • Streptomyces / drug effects
  • Streptomyces / genetics*
  • Streptomyces / metabolism
  • Streptomyces / physiology

Substances

  • Bacterial Proteins
  • Pyridones
  • Recombinant Fusion Proteins
  • Peptide Elongation Factor Tu
  • mocimycin

Associated data

  • GENBANK/X67057
  • GENBANK/X67058
  • GENBANK/X67059