Regulation of gene expression in Escherichia coli by the local anesthetic procaine

Granett, Sandra; Villarejo, Merna

doi:10.1016/0022-2836(82)90181-4

Cited by 20 publications

(22 citation statements)

References 12 publications

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“…Strains carrying the envZII allele are deficient in the synthesis of amylomaltase, maltose-binding protein, and the LamB porin (35). For LamB and maltosebinding protein, the effect appears to be at the transcriptional level (10,35 constitutive, positive regulator MalT. The regulation of malT is shown in Table 2.…”

Section: Resultsmentioning

confidence: 99%

Contrasting mechanisms of envZ control of mal and pho regulon genes in Escherichia coli

Case¹,

Bukau²,

Granett³

et al. 1986

J Bacteriol

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The envZII missense mutation in the regulatory gene envZ pleiotropically repressed synthesis of OmpF, alkaline phosphatase, and several proteins of the maltose regulon. Procaine treatment of wild-type cells resulted in the same phenotype through an envZ+-mediated mechanism. Here we show that envZII-procaine act differently on the mal and pho regulons. In the mal system, the expression of the positive regulator gene malT, measured as 0-galactosidase activity of a malT-lac+ operon fusion, was drastically reduced by procaine treatment or by the envZll mutation. In contrast, expression of the positive regulator of the pho regulon phoB was not reduced by procaine treatment. The products of the regulatory genes phoM, phoR, and phoU were also not required for procaine action. Procaine and envZII inhibited expression of only two products of the pho regulon, alkaline phosphatase and the PhoE porin. The conclusion that envZll-procaine act differently on the mal and the pho regulons is supported by our ability to isolate second-site mutations with a Mal' PhoAphenotype in an envZII strain.

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Section: Resultsmentioning

confidence: 99%

Contrasting mechanisms of envZ control of mal and pho regulon genes in Escherichia coli

Case¹,

Bukau²,

Granett³

et al. 1986

J Bacteriol

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“…It is known that growth of E. coli in the presence of procaine strongly reduces OmpF and increases OmpC levels in the outer membrane (15,35). These effects were shown to be mediated by EnvZ and OmpR (47,52) and were reported to be due to changes in transcription (38,47).…”

Section: Discussionmentioning

confidence: 99%

“…These proteins are encoded by the unlinked ompC and ompF genes (42,50). While the structural and functional properties of OmpC and OmpF are similar (22), their expression is regulated in opposite directions by a large number of environmental factors including medium osmolarity (51), temperature (24), pH (18), and the presence of compounds, such as procaine, that act as membrane perturbants (15,35,47). In the most thoroughly analyzed of these processes, the reciprocal osmoregulation of OmpC and OmpF is thought to primarily involve transcriptional control mediated by OmpR and EnvZ, the protein products of the ompB operon (16,17).…”

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confidence: 99%

micF antisense RNA has a major role in osmoregulation of OmpF in Escherichia coli

1994

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micF RNA, produced from a multicopy plasmid, was originally shown to be a major factor in negative osmoregulation of the OmpF outer membrane protein in Escherichia coli. However, subsequent experiments with a micF deletion strain suggested that chromosomal micF RNA was not a key component in this process. We report here that micF RNA is essential for the reduction in OmpF levels in cells grown in media of low-to-intermediate levels of osmolarity. Under these conditions, the amount of OmpF was reduced up to 60% in the parent strain while OmpF levels were not altered in the micF deletion mutant. In medium of higher osmolarity, OmpF synthesis was strongly inhibited in both strains. RNA measurements showed that micF RNA levels rose rapidly in cells grown in low-to-intermediate levels of osmolarity concomitant with the reduction in OmpF protein, while ompF mRNA decreased strongly only during high-osmolarity conditions. Taken together, these results strongly suggest that the negative osmoregulation of OmpF at low-to-intermediate osmolarity levels requires micF RNA and that this is masked at higher osmolarity by the known strong inhibition of OmpF transcription by OmpR. Results consistent with this model were also obtained by using procaine, a compound reported to inhibit ompF expression by a mechanism very similar to that involved in osmoregulation.

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“…The amounts of these proteins reciprocally vary depending upon external conditions such as medium osmolarity (50), carbon source (42) pH (25), or the presence of membrane perturbants (21,22,37,39).…”

mentioning

confidence: 99%

Procaine, a local anesthetic, signals through the EnvZ receptor to change the DNA binding affinity of the transcriptional activator protein OmpR

Rampersaud

Inouye

1991

J Bacteriol

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Local anesthetics are known to reduce the level of OmpF and increase the synthesis of OmpC in the outer membrane of Escherichia coli K-12. It has been shown that the anesthetics procaine and phenethyl alcohol (PEA) act at the transcriptional level for ompF and ompC and that in the case of procaine, its action is dependent on EnvZ, the membrane-bound signal transducer required for ompF and ompC expression. In an elfort to further understand how anesthetics regulate ompF and ompC expression, we have analyzed the DNA binding properties of OmpR (the transcriptional activator protein for ompF and ompC genes) from cells treated with procaine or PEA. Treatment of a wild-type cell with either anesthetic converted OmpR from a low-affinity DNA binding form to a high-affinity DNA binding form. The change in DNA binding affinity was correlated with alterations in outer membrane porin profiles and could occur in the absence of protein synthesis. A strain lacking EnvZ was unable to respond to procaine to produce either the shift in the OmpR DNA binding property or cause any change in the outer membrane porin profile. PEA treatment was also dependent on EnvZ for the alteration in the OmpR DNA binding property, but it could induce ompC expression in the absence of EnvZ.Further studies suggest that the amino-terminal region of EnvZ is responsible for the procaine signalling. Our results indicate that procaine and PEA regulate ompF and ompC expression by modifying the DNA binding properties of OmpR through EnvZ signal transduction.

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Regulation of gene expression in Escherichia coli by the local anesthetic procaine

Cited by 20 publications

References 12 publications

Contrasting mechanisms of envZ control of mal and pho regulon genes in Escherichia coli

Contrasting mechanisms of envZ control of mal and pho regulon genes in Escherichia coli

micF antisense RNA has a major role in osmoregulation of OmpF in Escherichia coli

Procaine, a local anesthetic, signals through the EnvZ receptor to change the DNA binding affinity of the transcriptional activator protein OmpR

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