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Papers In Press, published online ahead of print May 4, 2001
Department of Food Science and Technology, Seoul National University, Suwon 441-744
Corresponding Author: sangryu{at}snu.ac.kr
Mlc is a global regulator of carbohydrate metabolism. Recent studies have revealed that Mlc is depressed by protein-protein interaction with enzyme IICBGlc, a glucose-specific permease, which is encoded by ptsG. The mlc gene has been previously known to be transcribed by two promoters, P1 (+1) and P2 (+13), and have a binding site of its own gene product at +16. However, the mechanism of transcriptional regulation of the gene has not yet been established. In vitro transcription assays of the mlc gene showed that P2 promoter could be recognized by RNA polymerase containing the heat shock sigma factor
J. Biol. Chem, 10.1074/jbc.M101757200
Submitted on February 26, 2001
Revised on April 17, 2001
Accepted on May 4, 2001
Heat shock RNA polymerase (E
32) is involved in the transcription of mlc and crucial for induction of the Mlc regulon by glucose in Escherichia coli
32 (E32) as well as E70, while P1 promoter is only recognized by E70. The cyclic AMP receptor protein and cyclic AMP complex (CRP¡¤cAMP) increased expression from P2 but showed negative effect on transcription from P1 by E70, although it had little effect on transcription from P2 by E32 in vitro. Purified Mlc repressed transcription from both promoters, but with different degrees of inhibition. In vivo transcription assays using wild type and mlc strains indicated that the level of mlc expression was modulated less than two-fold by glucose in the medium with concerted action of CRP¡¤cAMP and Mlc. A dramatic increase in mlc expression was observed upon heat shock or in cells overexpressing 32, confirming that E32 is involved in the expression of mlc. Induction of ptsG and pts P0 transcription by glucose was also strictly dependent on E32. These results indicate that E32 plays an important role in balancing the relative concentration of Mlc and EIICBGlc in response to availability of glucose in order to maintain inducibility of the Mlc regulon at high growth temperature.
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