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
复制标题
DOI：
10.1016/s0378-1097(01)00182-3
复制
发表时间：
2001-06-12
影响因子：
2.1
通讯作者：
Yamane, T
中科院分区：
文献类型：
--
作者：
Maehara, A;Doi, Y;Yamane, T
研究了在反硝化副球菌聚羟基烷酸合成位点（phaZCPR）中发现的一种可能的调节蛋白PhaR。经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，从重组大肠杆菌中纯化的PhaR蛋白估计为22 kDa，与核苷酸序列计算的质量一致。通过尺寸排除色谱法测定其分子量为93 kDa，表明该蛋白为四聚体。凝胶迁移试验表明，PhaR特异性结合到phaC-phaP的基因间区。在使用大肠杆菌S30提取物的无细胞蛋白合成系统中，添加纯化的PhaR抑制了phaP基因的表达，这些结果表明PhaR是一种dna结合蛋白，可能在调控phaP基因表达中发挥作用(C) 2001出版于Elsevier Science B.V.，代表欧洲微生物学会联合会。
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 shirt 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 (C) 2001 Published by Elsevier Science B.V. on behalf of the Federation of European Microbiological Societies.