Fine-tuning in regulation of Clp protein content in Bacillus subtilis
Fine-tuning in regulation of Clp protein content in Bacillus subtilis
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DOI：
10.1128/jb.186.1.179-191.2004
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发表时间：
2004-01-01
影响因子：
3.2
通讯作者：
Hecker, M
中科院分区：
文献类型：
--
作者：
Gerth, U;Kirstein, J;Hecker, M
CLP控制的蛋白水解在枯草芽孢杆菌似乎发挥了重要作用，特别是在应力条件下。使用校准的蛋白质印迹分析来估计单个细胞内热诱导Clp分子的近似数目。根据这些数字，不同的Clp ATP酶似乎不竞争蛋白水解亚基ClpP。免疫共沉淀实验揭示了预测的特异性ClpX-ClpP、ClpC-ClpP和ClpE-ClpP相互作用。ClpE和ClpX在永久性热应激期间在野生型细胞中迅速降解，但在clpP突变体中几乎保持稳定，表明ClpP依赖性降解。特别是，ClpCP似乎参与了短寿命的ClpE ATP酶的降解，表明在翻译后水平上这种特定Clp ATP酶的负“自动调节”回路。在指数生长和热休克过程中的应力诱导的clp mRNA的半衰期的分析揭示了精确的调节每个Clp蛋白的合成在转录后水平，以及满足B的需要。枯草杆菌。
Clp-controlled proteolysis in Bacillus subtilis seems to play a substantial role, particularly under stress conditions. Calibrated Western blot analyses were used to estimate the approximate numbers of heat-inducible Clp molecules within a single cell. According to these numbers, the different Clp ATPases do not seem to compete for the proteolytic subunit ClpP. Coimmunoprecipitation experiments revealed the predicted specific ClpX-ClpP, ClpC-ClpP, and ClpE-ClpP interactions. ClpE and ClpX are rapidly degraded in wild-type cells during permanent heat stress but remained almost stable in a clpP mutant, suggesting ClpP-dependent degradation. In particular, ClpCP appeared to be involved in the degradation of the short-lived ClpE ATPase, indicating a negative "autoregulatory" circuit for this particular Clp ATPase at the posttranslational level. Analysis of the half-life of stress-inducible clp mRNAs during exponential growth and heat shock revealed precise regulation of the synthesis of each Clp protein at the posttranscriptional level as well to meet the needs of B. subtilis.