DNA sequence of the promoter region of the <i>ompC</i> gene and the amino acid sequence of the signal peptide of pro‐OmpC protein in <i>Escherichia coli</i>

Mizuno, Takeshi; Chou, Mei Yin; Inouye, Masayori

doi:10.1016/0014-5793(83)80364-0

Cited by 33 publications

(11 citation statements)

References 17 publications

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“…Picomolar to nanomolar concentrations of this 44-kDa protein were able to block attachment of S. typhimurium to macrophages. Partial amino acid sequence data obtained from the N-terminus of the 44-kDa protein completely matched a 15-mer of the OmpD of S. typhimurium SH 7454 [27] and the OmpC of E. coli K-12 [28]. A similar but not identical sequence is associated with the 36-kDa OmpC from S. typhimurium LT2 [18] and 14028 (see Table 2).…”

Section: -Kdamentioning

confidence: 73%

Macrophages recognize and adhere to an OmpD-like protein of Salmonella typhimurium

Negm

1998

FEMS Immunology and Medical Microbiology

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Murine peritoneal macrophages bind to Salmonella typhimurium in vitro in the absence of exogenous opsonins. We have identified an outer membrane protein of S. typhimurium that mediates this adhesion. Biotin-labeled macrophages were used to probe electroblotted envelope proteins of S. typhimurium that had been previously resolved by polyacrylamide electrophoresis under denaturing and reducing conditions. Macrophages bound to an outer membrane protein with an apparent molecular mass of 44 kDa. The protein was purified to homogeneity and free of detectable lipopolysaccharide. Limited microsequencing of this protein resulted in a 15-amino acid query sequence of A-E-V-Y-N-K-D-G-N-K-L-D-L-Y-G, which shares complete identity with a 15-mer of both the OmpD of S. typhimurium SH 7454 and the OmpC polypeptide of Escherichia coli K-12. Picomolar concentrations of this purified protein significantly inhibited the subsequent adherence of 35S-labeled S. typhimurium to macrophages in monolayers. We propose that this 44-kDa protein is involved in the recognition of S. typhimurium by macrophage during the initial stages of infection.

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Section: -Kdamentioning

confidence: 73%

Macrophages recognize and adhere to an OmpD-like protein of Salmonella typhimurium

Negm

1998

FEMS Immunology and Medical Microbiology

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“…The ompC and ompF genes have been cloned, and their total nucleotide sequences have been determined (11,20,21). The extensive homology between them in the coding DNA sequences and in the primary amino acid sequences is obvious (21).…”

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confidence: 99%

Characterization by deletion and localized mutagenesis in vitro of the promoter region of the Escherichia coli ompC gene and importance of the upstream DNA domain in positive regulation by the OmpR protein

Mizuno¹,

Mizushima²

1986

J Bacteriol

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The ompC gene codes for a major outer membrane protein whose expression is regulated by the ompR and envZ genes. Two sets of promoter deletion mutants, with upstream and downstream deletions, were constructed on a plasmid in vitro, and their promoter activity was studied by connecting them with the lacZ gene. The DNA sequence for the ompC promoter, including the -35 and -10 regions and the mRNA start site, was defined at the region about 100 base pairs upstream from the ATG initiation codon for the pro-OmpC protein. An additional 61-base-pair sequence extending upstream from the -35 region was required for the ompC promoter to function fully. After targeting the upstream region of the ompC promoter fused to the lacZ gene on a plasmid, in vitro-localized mutagenesis was performed to isolate cis-dominant mutations that affect ompC transcription. Four mutant groups, each of which had common phenotypes for expression and regulation of the gene, were identified. The individual groups also had common base substitutions. In two of the groups, the common base substitutions were localized in the upstream region of the ompC promoter, whereas in the other two they were localized in the -35 region. From these results, the upstream region of the ompC promoter was considered to be the domain responsible for activation by the ompR gene product.

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“…The sequence TCGAAG happened to be identical to the -35 sequence of deoP2 promoter (6). The three -10 sequences include GAGAAT from the OmpRdependent ompC promoter (8,9), TACTGT from the AraCdependent araBAD promoter (6,14), and TATACT from the constitutive Ipp P-5 promoter (16). The last -10 sequence has been shown to exert stronger transcriptional activities than the consensus -10 sequence (TATAAT; ref.…”

Section: Resultsmentioning

confidence: 99%

“…Because the sequences of these promoters diverge from the consensus sequence, RNA polymerase is either unable to bind to the promoters or to isomerize the closed complex of the promoter and RNA polymerase to the open complex without the aid of transcriptional activators. In the case of ompC and ompF, RNA polymerase is unable to transcribe these genes without OmpR because the -35 and -10 regions of these genes are quite different from the consensus sequence (7,8). The OmpR binding sites for both genes have been shown to exist in the regions from -40 to -100 (9,10).…”

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confidence: 99%

Enhancement of RNA polymerase binding to promoters by a transcriptional activator, OmpR, in Escherichia coli: its positive and negative effects on transcription.

Tsung

Brissette

Inouye

1990

Proc. Natl. Acad. Sci. U.S.A.

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The OmpR binding sequence (OBS) in the upstream region of the ompF promoter of Escherichia coli was fused to 27 synthetic promoters. Transcription from a number of weak promoters, regardless of their sequences, was dramatically activated in the presence of OmpR, a transcriptional activator. In vivo DNA footprinting revealed that OmpR enhanced the binding of RNA polymerase to the promoters. This enhancement was essential for transcription of weak promoters, while OmpR binding to the OBS fused to a strong promoter was inhibitory for transcription. These results indicate that OmpR stabilizes the formation of an RNA polymerasepromoter complex, possibly a closed promoter complex, and that a transcription activator can serve not only as a positive but also as a negative regulator for gene expression. There are a number of E. coli genes whose transcription is activated by a protein factor binding to a specific sequence upstream of the RNA polymerase recognition site (3-5). The RNA polymerase recognition sites for genes requiring transcriptional activators for "7O RNA polymerase are considerably different from the consensus sequence consisting of the -35 region (TTGACA) and the -10 region (TATAAT) relative to +1, the transcription initiation site (6). Because the sequences of these promoters diverge from the consensus sequence, RNA polymerase is either unable to bind to the promoters or to isomerize the closed complex of the promoter and RNA polymerase to the open complex without the aid of transcriptional activators. In the case of ompC and ompF, RNA polymerase is unable to transcribe these genes without OmpR because the -35 and -10 regions of these genes are quite different from the consensus sequence (7,8 Fig. LA) as well as the -35 and -10 promoter sequence. The lacZ gene coding sequence starting from the eighth codon is fused in-frame to the above ompF sequence. By site-specific mutagenesis the 7-base-pair (bp) DNA sequence immediately after the Cd box (Fig. lA) of ompF, TCACGGf (asterisk indicates the first base in the -35 region of ompF), was replaced with the sequence AAGAEI to generate a unique Bgl II site (underlined). The DNA sequence between this Bgl II site and an original Pst I site at the + 1 position (transcription initiation site) of ompF was then replaced with synthetic oligonucleotides. For example, the sequence for promoter 1 consists of the consensus -35 sequence (TTGACA), an 18-bp spacer sequence (CTrTAAGCTTCCGGCTCG), the -10 sequence of the ompC promoter (GAGAAT), and a 9-bp sequence (GICGACAAT) after the -10 sequence to generate a unique Sal I site (underlined). The spacer sequence is identical to that of the lac promoter (6) except that the sixth T residue (indicated by an asterisk) was changed to A to create a unique HindIII site (underlined). The 9-bp sequence after the -10 sequence is from the lacZ promoter sequence, where the A residue with an asterisk indicates the transcription initiation site. Other synthetic promoters (from no. 2 to no. 27) were constructed by replacing DNA seque...

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DNA sequence of the promoter region of the ompC gene and the amino acid sequence of the signal peptide of pro‐OmpC protein in Escherichia coli

Cited by 33 publications

References 17 publications

Macrophages recognize and adhere to an OmpD-like protein of Salmonella typhimurium

Macrophages recognize and adhere to an OmpD-like protein of Salmonella typhimurium

Characterization by deletion and localized mutagenesis in vitro of the promoter region of the Escherichia coli ompC gene and importance of the upstream DNA domain in positive regulation by the OmpR protein

Enhancement of RNA polymerase binding to promoters by a transcriptional activator, OmpR, in Escherichia coli: its positive and negative effects on transcription.

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