PhaR, a protein of unknown function conserved among short-chain-length polyhydroxyalkanoic acids producing bacteria, is a DNA-binding protein and represses Paracoccus denitrificans phaP expression in vitro.
PhaR, a protein of unknown function conserved among short-chain-length polyhydroxyalkanoic acids producing bacteria, is a DNA-binding protein and represses Paracoccus denitrificans phaP expression in vitro.
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DOI：
10.1111/j.1574-6968.2001.tb10685.x
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发表时间：
2001-06
影响因子：
2.1
通讯作者：
A. Maehara;Y. Doi;T. Nishiyama;Yasuo Takagi;S. Ueda;Hideo Nakano;Tsuneo Yamane
中科院分区：
文献类型：
--
作者：
A. Maehara;Y. Doi;T. Nishiyama;Yasuo Takagi;S. Ueda;Hideo Nakano;Tsuneo Yamane
研究人员对一种假定的调节蛋白 PhaR 进行了研究，该蛋白在脱氮副球菌的聚羟基链烷酸合成位点 (phaZCPR) 中被鉴定。重组大肠杆菌纯化的PhaR蛋白经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳测得分子量为22 kDa，与核苷酸序列计算的质量一致。通过尺寸排阻色谱法测定分子量为93 kDa，表明该蛋白质形成四聚体。凝胶迁移率变化分析表明 PhaR 特异性结合 phaC-phaP 的基因间区域。在使用大肠杆菌 S30 提取物的无细胞蛋白质合成系统中，通过添加纯化的 PhaR 来抑制 phaP 基因的表达。这些结果表明 PhaR 是一种 DNA 结合蛋白，可能在 phaP 基因表达的调节中发挥作用。
A putative regulatory protein, PhaR, which was identified in the polyhydroxyalkanoic acid synthetic locus (phaZCPR) in Paracoccus denitrificans, was investigated. The PhaR protein purified from a recombinant Escherichia coli was estimated to be 22 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, being consistent with the mass calculated from the nucleotide sequence. The molecular mass was determined to be 93 kDa by size-exclusion chromatography, suggesting that the protein formed a tetramer. A gel mobility shift assay showed that PhaR specifically bound to the intergenic region of phaC--phaP. In a cell-free protein synthesis system using E. coli S30 extract, the expression of the phaP gene was repressed by the addition of purified PhaR. These results suggest that PhaR is a DNA-binding protein and may play a role in the regulation of phaP gene expression.