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微阵列研究揭示了谷氨酸棒杆菌中依赖HrcA和HspR的系统对中度或重度热休克的“差异反应”。

Microarray studies reveal a 'differential response' to moderate or severe heat shock of the HrcA- and HspR-dependent systems in Corynebacterium glutamicum.

作者信息

Barreiro Carlos, Nakunst Diana, Hüser Andrea T, de Paz Héctor D, Kalinowski Jörn, Martín Juan F

机构信息

INBIOTEC (Instituto de Biotecnología de León), Parque Científico de León, Avda. Real, 1, 24006 León, Spain.

Institut für Genomforschung und Systembiologie, Centrum für Biotechnologie, Universität Bielefeld, Universitätsstrasse 25, 33615 Bielefeld, Germany.

出版信息

Microbiology (Reading). 2009 Feb;155(Pt 2):359-372. doi: 10.1099/mic.0.019299-0.

Abstract

Genome-wide transcription profile analysis of the heat-shocked wild-type strain under moderate (40 degrees C) and severe heat stress (50 degrees C) revealed that a large number of genes are differentially expressed after heat shock. Of these, 358 genes were upregulated and 420 were downregulated in response to moderate heat shock (40 degrees C) in Corynebacterium glutamicum. Our results confirmed the HrcA/controlling inverted repeat of chaperone expression (CIRCE)-dependent and HspR/HspR-associated inverted repeat (HAIR)-dependent upregulation of chaperones following heat shock. Other genes, including clusters of orthologous groups (COG) related to macromolecule biosynthesis and several transcriptional regulators (COG class K), were upregulated, explaining the large number of genes affected by heat shock. Mutants having deletions in the hrcA or hspR regulators were constructed, which allowed the complete identification of the genes controlled by those systems. The up- or downregulation of several genes observed in the microarray experiments was validated by Northern blot analyses and quantitative (real-time) reverse-transcription PCR. These analyses showed a heat-shock intensity-dependent response ('differential response') in the HspR/HAIR system, in contrast to the non-differential response shown by the HrcA/CIRCE-regulated genes.

摘要

对在中度(40摄氏度)和重度热应激(50摄氏度)下热激的野生型菌株进行全基因组转录谱分析,结果显示热激后大量基因存在差异表达。其中,谷氨酸棒杆菌在中度热激(40摄氏度)后,358个基因上调,420个基因下调。我们的结果证实了热激后伴侣蛋白表达受HrcA/伴侣蛋白表达控制反向重复序列(CIRCE)依赖性以及HspR/HspR相关反向重复序列(HAIR)依赖性上调。包括与大分子生物合成相关的直系同源簇(COG)和几个转录调节因子(COG分类K)在内的其他基因也上调,这解释了大量受热激影响的基因。构建了hrcA或hspR调节因子缺失的突变体,从而能够完全鉴定由这些系统控制的基因。通过Northern印迹分析和定量(实时)逆转录PCR验证了在微阵列实验中观察到的几个基因的上调或下调。这些分析表明,与HrcA/CIRCE调节基因显示的非差异反应相反,HspR/HAIR系统存在热激强度依赖性反应(“差异反应”)。

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