Elongation factor Tu1 of the antibiotic GE2270A producer Planobispora rosea has an unexpected resistance profile against EF-Tu targeted antibiotics.
Sensitivity of EF-Tu1 of the GE2270A producer Planobispora rosea towards GE2270A, pulvomycin and kirromycin was determined by band-shift assays for EF-Tu1-antibiotic complex formation and by in vitro translation experiments. EF-Tu1 of P. rosea appeared to be not only totally resistant to GE2270A, but also ten times more resistant to kirromycin than EF-Tu1 of Streptomyces coelicolor. In contrast, P. rosea EF-Tu1 was found to be not resistant to pulvomycin, an antibiotic that just like GE2270A blocks EF-Tu x GTP x aminoacyl-tRNA complex formation. Previous in vivo and in vitro experiments with mixed populations of antibiotic resistant and sensitive EF-Tu species had shown that sensitivity to kirromycin and pulvomycin is dominant over resistance. In the case of GE2270A we observed, however, that sensitivity is recessive to resistance, which again points to a different action mechanism than in the case of pulvomycin. Besides the tuf1 gene encoding the regular elongation factor EF-Tu1 a gene similar to S. coelicolor tuf3 for a specialized EF-Tu was located in the P. rosea genome. The tuf1 gene was isolated and sequenced. The amino acid sequence of EF-Tul of P. rosea not only exhibits an unusual Tyr160 substitution (comparable to those described for kirromycin-resistant EF-Tus), but also shows significant changes of conserved amino acids in domain 2 that may be responsible for GE2270A resistance (the latter do not resemble those leading to pulvomycin resistance). P. rosea EF-Tu1 thus is a first example of a bacterial EF-Tu with resistance against two divergently acting antibiotics.
Möhrle
V G
VG
Department of Biochemistry, Leiden Institute of Chemistry, Gorlaeus Laboratories, Leiden University, The Netherlands.
Tieleman
L N
LN
Kraal
B
B
D000192:Actinomycetales / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins; D002474:Cell-Free System; D003001:Cloning, Molecular; D004352:Drug Resistance, Microbial; D005798:Genes, Bacterial; D007700:Kinetics; D008969:Molecular Sequence Data; D010452:Peptide Biosynthesis; D010444:Peptide Elongation Factor Tu / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D010455:Peptides; D010456:Peptides, Cyclic / Q000494:pharmacology; D014176:Protein Biosynthesis; D017433:Protein Structure, Secondary; D011728:Pyridones / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D010455:Peptides; D010456:Peptides, Cyclic; D011728:Pyridones; D011994:Recombinant Proteins; D013844:Thiazoles; C012521:pulvomycin; C012001:polyphenylalanine; D010444:Peptide Elongation Factor Tu; C068122:GE 2270 A; C008536:mocimycin