Response regulator Spo0A phosphorylated from Bacillus subtilis is efficiently in Escherichia coli
Response regulator Spo0A phosphorylated from Bacillus subtilis is efficiently in Escherichia coli
复制标题
DOI：
10.1016/s0378-1097(03)00321-5
复制
发表时间：
2003-06-27
影响因子：
2.1
通讯作者：
Barák, I
中科院分区：
文献类型：
--
作者：
Ladds, JC;Muchová, K;Barák, I
双组分系统的反应调节蛋白介导了细菌对其不断变化的环境的许多适应。大多数反应调节因子是改变特定基因组转录水平的转录因子。反应调节因子的激活需要它们在一个保守的天冬氨酸残基上被同源的传感器激酶磷酸化。由于这个原因，在缺乏必要的传感器激酶的情况下，重组反应调节因子的表达预计会产生非活性状态下的非磷酸化产物。然而，对于枯草芽孢杆菌中控制产孢的反应调节因子Spo0A，我们发现纯化的重组蛋白的很大一部分被磷酸化。这种磷酸化的成分是二聚体，与DNA中的Spo0A识别序列结合。用spo0a特异性磷酸酶Spo0E处理会导致二聚体的分离和DNA结合的丧失。因此，有必要用同源磷酸酶预处理重组Spo0A制剂，以产生分子的完全失活状态。(C) 2003欧洲微生物学会联合会。Elsevier Science B.V.版权所有。
The response regulator proteins of two-component systems mediate many adaptations of bacteria to their ever-changing environment. Most response regulators are transcription factors that alter the level of transcription of specific sets of genes. Activation of response regulators requires their phosphorylation on a conserved aspartate residue by a cognate sensor kinase. For this reason, expression of a recombinant response regulator in the absence of the requisite sensor kinase is expected to yield an unphosphorylated product in the inactive state. For Spo0A, the response regulator controlling sporulation in Bacillus subtilis however, we have found that a significant fraction of the purified recombinant protein is phosphorylated. This phosphorylated component is dimeric and binds to Spo0A recognition sequences in DNA. Treatment with the Spo0A-specific phosphatase, Spo0E, leads to dissociation of the dimers and loss of DNA binding. It is therefore necessary to pre-treat recombinant Spo0A preparations with the cognate phosphatase, to generate the fully inactive state of the molecule. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.