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J. Biol. Chem., Vol. 279, Issue 32, 33253-33262, August 6, 2004

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

Hyoung Seok Kim, Tae Sun Kang, Joon Sik Hyun, and Hyen Sam Kang¶

From the Department of Microbiology, School of Biological Sciences, Seoul National University, San 56-1, Shillim-dong, Kwanak-gu, 151-742, Korea

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(N₂₇)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. (i) It has a native class III cAMP-receptor protein (CRP)-dependent promoter with two CRP-binding sites. (ii) The downstream CRP-binding site II has higher affinity. (iii) Binding of cAMP-CRP to both sites (I + II) is required for maximal expression. We have also shown that the PaaX-binding site overlaps with the CRP-binding site I. This implies that PaaX and the cAMP-CRP compete for binding to the region around the CRP-binding site I and therefore have antagonistic effects on pga expression.

Received for publication, April 20, 2004 , and in revised form, May 14, 2004.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains Data 1 and 2 and Tables A and B.

Present address: CJ Corp., R & D Center of Bioproducts, San 522-1, Dokpyong-Ri, Majang-Myon, Ichon-Si, Kyonggi-Do, 467-812, Korea.

Supported by BK21 Research Fellowship from the Ministry of Education and Human Resources Development.

^¶ To whom correspondence should be addressed. Tel.: 82-2-880-6701; Fax: 82-2-876-4440; E-mail: khslab{at}snu.ac.kr.

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