Novel mechanism of Escherichia coli porin regulation.
A novel mechanism of Escherichia coli porin regulation was discovered from multicopy suppressors that permitted growth of cells expressing a mutant OmpC protein in the absence of DegP. Analyses of two suppressors showed that both substantially lowered OmpC expression. Suppression activities were confined to a short DNA sequence, which we designated ipeX for inhibition of porin expression, and to DNA containing a 3'-truncated ompR gene. The major effect of ipeX on ompC expression was exerted posttranscriptionally, whereas the truncated OmpR protein reduced ompC transcription. ipeX was localized within an untranslated region of 247 base pairs between the stop codon of nmpC-a remnant porin gene from the cryptic phage qsr' (DLP12) genome-and its predicted Rho-independent transcriptional terminator. Interestingly, another prophage, PA-2, which encodes a porin similar to NmpC, known as Lc, has sequences downstream from lc identical to that of ipeX. PA-2 lysogenization leads to Lc expression and OmpC inhibition. Our data show that the synthesis of the lc transcript, whose 3' end contains the corresponding ipeX sequence, inhibits OmpC expression. Overexpression of ipeX RNA inhibited both OmpC and OmpF expression but not that of OmpA. ompC-phoA chimeric gene constructs revealed a 248-bp untranslated region of ompC required for ipeX-mediated inhibition. However, no sequence complementarity was found between ipeX and this region of ompC, indicating that inhibition may not involve simple base pairing between the two RNA molecules. The effect of ipeX on ompC, but not on ompF, was independent of the RNA chaperone Hfq.
Castillo-Keller
Maria
M
Faculty of Cellular and Molecular Sciences, School of Life Sciences, Arizona State University, Tempe, AZ 85287-4501, USA.
Vuong
Phu
P
Misra
Rajeev
R
D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D018388:Codon, Terminator; D003090:Coliphages / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D006360:Heat-Shock Proteins / Q000235:genetics; D009690:Nucleic Acid Conformation; D033903:Periplasmic Proteins / Q000235:genetics; D018272:Porins / Q000235:genetics* / Q000378:metabolism; D039002:Prophages / Q000502:physiology; D012329:RNA, Bacterial / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics; D013489:Suppression, Genetic; D015534:Trans-Activators / Q000235:genetics
D001426:Bacterial Proteins; D018388:Codon, Terminator; D006360:Heat-Shock Proteins; C113724:OmpC protein; D033903:Periplasmic Proteins; D018272:Porins; D012329:RNA, Bacterial; D015534:Trans-Activators; C112177:osmolarity response regulator proteins; C061682:DegP protease; D012697:Serine Endopeptidases