Isolation of Tn917 insertional mutants of Bacillus subtilis that are resistant to the protonophore carbonyl cyanide m-chlorophenylhydrazone.
Tn917 transposition libraries prepared from Bacillus subtilis were screened for mutants that had insertions in the chromosome resulting in resistance to the protonophore carbonylcyanide m-chlorophenylhydrazone (CCCP). Five such strains were characterized. Three of these were found to have distinct insertion sites that resulted in changes in fatty acid composition of the membrane lipids. The lipid changes were qualitatively similar to changes observed earlier in CCCP-resistant strains of B. subtilis that had been isolated after chemical mutagenesis. However, the extent of the changes was more modest, correlating with a lower level of protonophore-resistance. One of these mutants was disrupted in a gene homologous to the Escherichia coli rho gene, as reported earlier (Quirk et al. (1993) J. Bacteriol. 175, 647-654), one was disrupted in a new member of the two-component signalling systems, and the third was disrupted in a new gene of unknown function that apparently forms an operon with transporter genes. The other two CCCP-resistant mutants were disrupted in genes that are likely to encode membrane transporters; the disruption of these genes may have reduced the transmembrane ion leaks during growth, thus conferring modest protonophore-resistance. In one of these strains, the disrupted gene is part of an apparent operon that is a homologue of iron uptake operons from other prokaryotes.
Department of Biochemistry, Mount Sinai School of Medicine of the City University of New York, NY 10029.
P G
PG
Quirk
A A
AA
Guffanti
S
S
Clejan
J
J
Cheng
T A
TA
Krulwich
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology*; D017124:Conserved Sequence; D004251:DNA Transposable Elements; D004352:Drug Resistance, Microbial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005227:Fatty Acids / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics; D008563:Membrane Lipids / Q000235:genetics / Q000378:metabolism; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D016296:Mutagenesis; D016254:Mutagenesis, Insertional; D016366:Open Reading Frames; D012234:Rho Factor / Q000235:genetics; D017385:Sequence Homology; D001616:beta-Galactosidase / Q000235:genetics
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D005227:Fatty Acids; D008563:Membrane Lipids; D012234:Rho Factor; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D001616:beta-Galactosidase; D019204:GTP-Binding Proteins