Characterization of the regulatory region of the Escherichia coli penicillin acylase structural gene

Valle, Fernando; Gosset, Guillermo; Tenório, Bruno Mendes; Oliver, G.; Bolívar, Francisco

doi:10.1016/0378-1119(86)90316-1

Cited by 33 publications

(20 citation statements)

References 14 publications

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“…In spite of the high similarity of the pac genes from E. coli W ATCC 11105 and Kluyvera citrophila, the pac promoter of K. citrophila was located overlapping the sequence of the CRP2 binding site [12], this is, far upstream from the promoter (P3 promoter) proposed for the pac gene of E. coli W ATCC 11105 [6] (Fig. 1A).…”

Section: Analysis Of Pac Promotermentioning

confidence: 99%

“…1B) was inducible by PAA, whereas E. coli HB101 (pAJ19) produced PGA constitutively. Sequencing of plasmid pAJ19 [11] revealed that the cloned fragment only contains the ¢rst 53 bp of the 5P non-coding region of the pac gene that exclusively harbors the 310 box of the previously proposed pac promoter [6] (Fig. 1B, hereafter named P3 promoter).…”

Section: Expression Of the Pac Gene In E Coli K12 Strainsmentioning

confidence: 99%

“…This ¢nding pointed to the possibility that other promoters might be involved in the expression of the E. coli pac gene. To investigate this possibility, we constructed plasmid pRS55E that contains a lacZ gene fused to a fragment lacking the 310 box of the previously proposed pac promoter [6] or P3 promoter (Fig. 1B).…”

Section: Analysis Of Pac Promotermentioning

confidence: 99%

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New insights into the regulation of the pac gene from Escherichia coli W ATCC 11105

Roa

1999

FEMS Microbiology Letters

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The regulation of the pac gene encoding the penicillin G acylase of Escherichia coli W ATCC 11015 has been investigated by a molecular approach using lacZ as a reporter gene. This analysis revealed that a region of 170 bp located upstream of the pac structural gene contains the regulatory sequences that control its expression. The cAMP receptor protein is involved not only in the catabolite repression of penicillin G acylase production caused by glucose but also in the induction of pac gene expression by phenylacetic acid. Primer extension analyses have demonstrated that the transcription of the pac gene can be initiated from at least three different promoters. Although all these promoters are functional, their relative activity depends on the transcribed gene, the P1 and P3 promoters being more active in the presence of the pac gene, whereas the P2 promoter was stronger when the upstream region of the pac gene was fused to the lacZ reporter. A deletion of the region surrounding the 310 box of the P3 promoter produced a constitutive expression of the fused gene indicating that this sequence is required for phenylacetic acid induction and suggesting that the expression of the pac gene is regulated by a repression mechanism. This work reveals that the regulation of the pac gene is more complex than previously envisioned and provides new clues to investigate further this interesting regulatory system. ß

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Section: Analysis Of Pac Promotermentioning

confidence: 99%

Section: Expression Of the Pac Gene In E Coli K12 Strainsmentioning

confidence: 99%

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New insights into the regulation of the pac gene from Escherichia coli W ATCC 11105

Roa

1999

FEMS Microbiology Letters

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“…Previous studies of the E. coli pga promoter identified two putative CRP-binding sites (14,34) and two possible IHFbinding sites (21); however, there is no direct evidence that CRP or IHF proteins bind to the promoter (35). It has been inferred that the pga promoter is a class III CRP-dependent promoter (21).…”

mentioning

confidence: 99%

Regulation of Penicillin G Acylase Gene Expression in Escherichia coli by Repressor PaaX and the cAMP-cAMP Receptor Protein Complex

Kim¹,

Kang²,

Hyun³

et al. 2004

Journal of Biological Chemistry

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The pga gene of Escherichia coli W ATCC11105 encodes a penicillin G acylase whose expression is regulated at both the transcriptional and post-transcriptional level. In this work we have shown that PaaX is the repressor of pga expression, and we have identified its binding consensus as TGATTC(N 27 )GAATCA. We conclude that the process of "PAA induction" actually involves relief of pga from repression by PaaX. Other features of the pga promoter have also been characterized.

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“…Periplasmic processing steps consist of autoproteolysis (␣ϩ␤) of the precursor and folding of the mature PAC. (27,29,32), but direct evidence of the binding of these proteins to the Ppac promoter has not been obtained. Using the PPAC fragment as a probe, we tested the binding of purified CRP (kindly provided by A. Kolb) and IHF (kindly provided by F. Boccard) to the Ppac region by use of a gel retardation assay (Fig.…”

mentioning

confidence: 99%

The PaaX Repressor, a Link between Penicillin G Acylase and the Phenylacetyl-Coenzyme A Catabolon of Escherichia coli W

2004

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The pac gene, encoding the penicillin G acylase from Escherichia coli W, is regulated by the PaaX repressor of the phenylacetate catabolic pathway. pac expression depends on the synthesis of phenylacetyl-coenzyme A. PaaX and the cyclic AMP receptor protein (CRP) bind in vitro to the Ppac promoter region. A palindromic sequence proposed as the PaaX operator is located upstream of the ؊35 box overlapping a CRP binding site, an unusual position that suggests a novel regulatory mechanism.

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Characterization of the regulatory region of the Escherichia coli penicillin acylase structural gene

Cited by 33 publications

References 14 publications

New insights into the regulation of the pac gene from Escherichia coli W ATCC 11105

New insights into the regulation of the pac gene from Escherichia coli W ATCC 11105

Regulation of Penicillin G Acylase Gene Expression in Escherichia coli by Repressor PaaX and the cAMP-cAMP Receptor Protein Complex

The PaaX Repressor, a Link between Penicillin G Acylase and the Phenylacetyl-Coenzyme A Catabolon of Escherichia coli W

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