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Volume 150, Issue 3

Redox property and regulation of PpsR, a transcriptional repressor of photosystem gene expression in Free

Abstract

PpsR from is involved in the repression of photosystem gene expression. The PpsR protein was heterologously overexpressed and purified to homogeneity. Gel mobility shift assay showed that the purified PpsR has DNA-binding activity. SDS-PAGE analysis showed that some portions of PpsR were oxidized, indicating that intramolecular or intermolecular disulphide bonds were formed between the two cysteines in each subunit. When the disulphide bond of PpsR was reduced by DTT, the binding activity of PpsR to the promoter region distinctly increased. The changes in protein level and DNA-binding activity of PpsR were observed in a conjugant with an extra copy of the gene and in a PpsR-null mutant (PPS1), respectively. Both cysteines in PpsR existed in their reduced form under aerobic, anaerobic-dark and anaerobic-light growth conditions, as determined using thiol-specific chemical modification. In an AppA-null mutant (APP11), the binding activity and the amount of PpsR decreased compared to those of the wild-type and an -complemented strain, and decreased even more under anaerobic-dark conditions than under aerobic conditions. PpsR had a redox-sensitive property but retained its reduced state in the cell, and its amount was reduced by disruption of AppA.

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Keyword(s): GMSA, gel mobility shift assay and NEM, N-ethylmaleimide
SGM
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2004-03-01
2024-04-19
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Redox property and regulation of PpsR, a transcriptional repressor of photosystem gene expression in Rhodobacter sphaeroides
Microbiology 150, 697 (2004); https://doi.org/10.1099/mic.0.26777-0
/content/journal/micro/10.1099/mic.0.26777-0
/content/journal/micro/10.1099/mic.0.26777-0
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