Unbalanced membrane phospholipid compositions affect transcriptional expression of certain regulatory genes in Escherichia coli.
The amount of porin protein OmpF in the outer membrane of Escherichia coli was reduced to one-third by the pgsA3 mutation that diminishes the amount of phosphatidylglycerol and cardiolipin in the membrane, whereas a cls (cardiolipin synthase) null mutation had no effect. Osmoregulation of OmpF was functional in the pgsA3 mutant. As assessed by the beta-galactosidase activities of lacZ fusions, the ompF expression was not reduced at the transcriptional level but was reduced about threefold at the posttranscriptional level by pgsA3. This reduction was mostly restored by a micF null mutation, and the micF RNA that inhibits the ompF mRNA translation was present 1.3 to 1.4 times more in the pgsA3 mutant, as assayed by RNase protection and Northern blot analyses. Elevation of the level of micF RNA was not restricted to acidic-phospholipid deficiency: OmpF was hardly detected and micF RNA was present 2.7 to 2.8 times more in a pssA null mutant that lacked phosphatidylethanolamine. Other common phenotypes of pgsA3 and pssA null mutants, reduced rates of cell growth and phospholipid synthesis, were not the cause of micF activation. Salicylate, which activates micF expression and inhibits cell motility, did not repress the flagellar master operon. These results imply that an unbalanced phospholipid composition, rather than a decrease or increase in the amount of specific phospholipid species, induces a phospholipid-specific stress signal to which certain regulatory genes respond positively or negatively according to their intrinsic mechanisms.
Inoue
K
K
Department of Biochemistry and Molecular Biology, Saitama University, Urawa, Japan.
Matsuzaki
H
H
Matsumoto
K
K
Shibuya
I
I
D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D010582:Bacteriophage lambda; D002462:Cell Membrane / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics / Q000502:physiology*; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005809:Genes, Regulator; D005838:Genotype; D008563:Membrane Lipids / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014158:Transcription, Genetic; D017855:Transferases (Other Substituted Phosphate Groups) / Q000096:biosynthesis / Q000235:genetics*; D001616:beta-Galactosidase / Q000096:biosynthesis
D001425:Bacterial Outer Membrane Proteins; D008563:Membrane Lipids; D010743:Phospholipids; D011993:Recombinant Fusion Proteins; D017855:Transferases (Other Substituted Phosphate Groups); C020676:CDP-diacylglycerol-glycerol-3-phosphate 3-phosphatidyltransferase; D001616:beta-Galactosidase