Some features of DNA-binding proteins involved in the regulation of theStreptomyces aureofaciens gap gene, encoding glyceraldehyde-3-phosphate dehydrogenase
Tóm tắt
AgapR gene, encoding a protein similar to the AraC/XylS family of bacterial transcriptional regulators, was previously identified upstream of thegap gene, coding for glyceraldehyde-3-phosphate dehydrogenase inStreptomyces aureofaciens. The GapR protein overproduced inEscherichia coli was shown to bind to thegap-P promoter region. Using the gel mobility shift assay with cell-free protein extracts from different developmental stages ofS. aureofaciens, we identified several other proteins, in addition to GapR, that specifically bound to theS. aureofaciens gap-P promoter region. When cell-free extracts fromS. aureofaciens cultivated in liquid medium with glucose were analyzed, only one complex corresponding to GapR was detected. A new protein interacting with thegap-P promoter was detected in stationary culture ofS. aureofaciens grown in the presence of mannitol as carbon source. The GapR protein was partially purified fromS. aureofaciens cultivated in liquid medium containing glucose and used for binding studies. DNA footprinting analysis revealed an identical protected region as previously identified for the GapR protein overproduced fromEscherichia coli. The direct role of the GapR protein in the regulation ofgap expression inS. aureofaciens in vivo was confirmed but regulation ofgap expression seems to be more complex, possibly involving other regulatory protein(s), depending on the developmental stage ofS. aureofaciens.
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