Avedissian M, Lessing D, Gober J W, Shapiro L, Gomes S L
Departamento de Bioquímica, Universidade de São Paulo, Brazil.
J Bacteriol. 1995 Jun;177(12):3479-84. doi: 10.1128/jb.177.12.3479-3484.1995.
The bacterial heat shock proteins DnaK and DnaJ are members of a class of molecular chaperones that are required for a wide variety of cellular functions at normal growth temperatures. In Caulobacter crescentus, the expression of the dnaKJ operon is regulated both temporally during the normal cell cycle and by heat shock. Analysis of deletions and base substitutions in the 5' region of the operon established the presence of two functional promoters: a heat shock-inducible promoter, P1, with characteristics of a sigma 32 promoter, and an adjacent sigma 70-like promoter, P2. Transcription initiating at the sigma 70-like promoter is under strict temporal control, whereas transcription initiating at the heat shock promoter at 30 degrees C is not. Transcription of dnaKJ occurs during a short period in the cell cycle, concomitant with the onset of DNA replication. Deletions in the 5' region have also revealed that all cis-acting sites required for temporal control of transcription reside within 50 bases of the P2 start site. Transcripts initiating from either the P1 or the P2 promoter have an RNA leader sequence with a high probability of forming an extensive secondary structure. Deletion of this leader sequence resulted in an increased rate of expression in both transcriptional and translational fusions. Although the temporal control of expression at physiological temperatures is not affected by the presence or absence of the leader sequence, changes in mRNA secondary structure may contribute to the modulation of DnaK and DnaJ levels at normal temperatures and during heat shock.
细菌热休克蛋白DnaK和DnaJ是一类分子伴侣的成员,它们在正常生长温度下参与多种细胞功能。在新月柄杆菌中,dnaKJ操纵子的表达在正常细胞周期中受到时间调控,同时也受热休克调控。对该操纵子5'区域的缺失和碱基替换分析确定了两个功能启动子的存在:一个热休克诱导型启动子P1,具有σ32启动子的特征,以及一个相邻的类σ70启动子P2。从类σ70启动子起始的转录受到严格的时间控制,而在30℃时从热休克启动子起始的转录则不受此控制。dnaKJ的转录在细胞周期的短时间内发生,与DNA复制的开始同步。5'区域的缺失还表明,转录时间控制所需的所有顺式作用位点都位于P2起始位点的50个碱基范围内。从P1或P2启动子起始的转录本都有一个RNA前导序列,很可能形成广泛的二级结构。删除该前导序列导致转录和翻译融合中的表达速率增加。尽管生理温度下表达的时间控制不受前导序列存在与否的影响,但mRNA二级结构的变化可能有助于在正常温度和热休克期间调节DnaK和DnaJ的水平。