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J Biol Chem, Vol. 274, Issue 36, 25398-25402, September 3, 1999
From the Products of the pts operon of
Escherichia coli have multiple physiological roles such as
sugar transport, and the operon is controlled by two promoters, P0 and
P1. Expression of the pts P0 promoter that is increased
during growth in the presence of glucose is also activated by cAMP
receptor protein·cAMP. Based on the existence of a sequence that has
a high similarity with the known Mlc binding site in the promoter, the
effects of the Mlc protein on the pts P0 promoter
expression were studied. In vivo transcription assays using
wild type and mlc-negative E. coli strains
grown in the presence and absence of glucose indicate that Mlc
negatively regulates expression of the P0 promoter, and Mlc-dependent repression is relieved by glucose in the
growth medium. In vitro transcription assay using purified
recombinant Mlc showed that Mlc repressed transcription from the P0 but
did not affect the activity of the P1. DNase I footprinting experiments revealed that a Mlc binding site was located around +1 to +25 of the
promoter and that Mlc inhibited the binding of RNA polymerase to the P0
promoter. Cells overexpressing Mlc showed a very slow fermentation rate
compared with the wild type when grown in the presence of various
phosphoenolpyruvate-carbohydrate phosphotransferase system
sugars but few differences in the presence of
non-phosphoenolpyruvate-carbohydrate phosphotransferase system sugars
except maltose. These results suggest that the pts operon
is one of major targets for the negative regulation by Mlc, and thus
Mlc regulates the utilization of various sugars as well as glucose in
E. coli. The possibility that the inducer of Mlc may not be
sugar or its derivative but an unknown factor is proposed to explain
the Mlc induction mechanism by various sugars.
Purification of Mlc and Analysis of Its Effects on the
pts Expression in Escherichia coli
,,
Department of Microbiology, College of
Medicine, Chungbuk National University, Chongju, 361-763 Korea and
the ¶ Department of Microbiology, Seoul National University,
Seoul, 151-742 Korea
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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