PMID:7928983
| Citation |
Stauffer, LT and Stauffer, GV (1994) Characterization of the gcv control region from Escherichia coli. J. Bacteriol. 176:6159-64 |
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| Abstract |
We constructed a set of deletions upstream of the gcv promoter and analyzed the effects of the deletions on expression of a gcvT-lacZ gene fusion. A deletion that ends at position -313 upstream of the transcription initiation site (+1) results in reduced levels of gcvT-lacZ expression, but the fusion is still inducible by glycine and repressible by purines. A deletion that ends at position -169 results in loss of both GcvA- and Lrp-mediated activation of the gcvT-lacZ fusion. The endpoints of delta -313 and delta -169 also define a site that down-regulates gcvT-lacZ expression two- to threefold. A deletion that ends at position -89 upstream from the transcription initiation site still shows PurR-mediated repression, suggesting that PurR-mediated repression is not by direct interference with the GcvA- and Lrp-mediated regulatory mechanism(s). Gel mobility shift assays and DNase I footprinting showed that Lrp protein binds to multiple sites upstream of the gcv promoter, from about bp -92 to bp -229. The results suggest that the gcv regulatory region is complex, with numerous cis-acting sites that are required for normal gcv expression. |
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| Keywords |
Amino Acid Oxidoreductases/biosynthesis; Amino Acid Oxidoreductases/genetics; Aminomethyltransferase; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Base Sequence; Carrier Proteins/biosynthesis; Carrier Proteins/genetics; DNA Mutational Analysis; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; Escherichia coli/genetics; Escherichia coli Proteins; Gene Expression Regulation, Enzymologic; Glycine/metabolism; Hydroxymethyl and Formyl Transferases; Leucine-Responsive Regulatory Protein; Molecular Sequence Data; Multienzyme Complexes/biosynthesis; Multienzyme Complexes/genetics; Recombinant Fusion Proteins/biosynthesis; Regulatory Sequences, Nucleic Acid/genetics; Sequence Deletion; Transcription Factors; Transferases/biosynthesis; Transferases/genetics |
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