Liberek K, Georgopoulos C
Departement de Biochimie Medicale, Centre Medical Universitaire, Genève, Switzerland.
Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11019-23. doi: 10.1073/pnas.90.23.11019.
All organisms respond to various forms of stress, including heat shock. The heat shock response has been universally conserved from bacteria to humans. In Escherichia coli the heat shock response is under the positive transcriptional control of the sigma 32 polypeptide and involves transient acceleration in the rate of synthesis of a few dozen genes. Three of the heat shock genes--dnaK, dnaJ, and grpE--are special because mutations in any one of these lead to constitutive levels of heat shock gene expression, implying that their products negatively autoregulate their own synthesis. The DnaK, DnaJ, and GrpE proteins have been known to function in various biological situations, including bacteriophage lambda replication. Here, we report the formation of an ATP hydrolysis-dependent complex of DnaJ, sigma 32, and DnaK proteins in vitro. This DnaJ-sigma 32-DnaK complex has been seen under different conditions, including glycerol gradient sedimentation and co-immunoprecipitation. The DnaK and DnaJ proteins in the presence of ATP can interfere with the efficient binding of sigma 32 to the RNA polymerase core, and are capable of disrupting a preexisting sigma 32-RNA polymerase complex. Our results suggest a possible mechanism for the autoregulation of the heat shock response.
所有生物体都会对包括热休克在内的各种形式的应激作出反应。从细菌到人类,热休克反应一直普遍存在。在大肠杆菌中,热休克反应受σ32多肽的正向转录调控,涉及几十种基因合成速率的短暂加速。其中三个热休克基因——dnaK、dnaJ和grpE——很特别,因为这三个基因中任何一个发生突变都会导致热休克基因表达达到组成型水平,这意味着它们的产物会对自身的合成进行负向自我调节。已知DnaK、DnaJ和GrpE蛋白在包括噬菌体λ复制在内的各种生物学过程中发挥作用。在此,我们报告了在体外形成了一种依赖ATP水解的DnaJ、σ32和DnaK蛋白复合物。在不同条件下,包括甘油梯度沉降和共免疫沉淀,都观察到了这种DnaJ-σ32-DnaK复合物。在ATP存在的情况下,DnaK和DnaJ蛋白会干扰σ32与RNA聚合酶核心的有效结合,并且能够破坏预先存在的σ32-RNA聚合酶复合物。我们的结果提示了热休克反应自我调节的一种可能机制。