National Institute for Basic Biology (NIBB), Okazaki 444-8585, Japan.
Department of Plant Sciences, School of Life Sciences, University of Hyderabad, Hyderabad 500 046, India.
Microbiology (Reading). 2010 Feb;156(Pt 2):442-451. doi: 10.1099/mic.0.031823-0. Epub 2009 Nov 19.
The crhR gene for RNA helicase, CrhR, was one of the most highly induced genes when the cyanobacterium Synechocystis sp. PCC 6803 was exposed to a downward shift in ambient temperature. Although CrhR may be involved in the acclimatization of cyanobacterial cells to low-temperature environments, its functional role during the acclimatization is not known. In the present study, we mutated the crhR gene by replacement with a spectinomycin-resistance gene cassette. The resultant DeltacrhR mutant exhibited a phenotype of slow growth at low temperatures. DNA microarray analysis of the genome-wide expression of genes, and Northern and Western blotting analyses indicated that mutation of the crhR gene repressed the low-temperature-inducible expression of heat-shock genes groEL1 and groEL2, at the transcript and protein levels. The kinetics of the groESL co-transcript and the groEL2 transcript after addition of rifampicin suggested that CrhR stabilized these transcripts at an early phase, namely 5-60 min, during acclimatization to low temperatures, and enhanced the transcription of these genes at a later time, namely 3-5 h. Our results suggest that CrhR regulates the low-temperature-inducible expression of these heat-shock proteins, which, in turn, may be essential for acclimatization of Synechocystis cells to low temperatures.
当蓝藻集胞藻 PCC 6803 暴露于环境温度下降时,crhR 基因(RNA 解旋酶 CrhR 的基因)是最被高度诱导的基因之一。虽然 CrhR 可能参与了蓝藻细胞对低温环境的适应,但它在适应过程中的功能作用尚不清楚。在本研究中,我们通过替换带有壮观霉素抗性基因盒的方法突变了 crhR 基因。所得的 DeltacrhR 突变体表现出低温生长缓慢的表型。对全基因组基因表达的 DNA 微阵列分析以及 Northern 和 Western 印迹分析表明,crhR 基因突变抑制了热休克基因 groEL1 和 groEL2 在低温诱导下的转录和蛋白水平表达。在加入利福平后 groESL 共转录物和 groEL2 转录物的动力学表明,CrhR 在低温适应的早期阶段(5-60 分钟)稳定这些转录物,并在稍后的时间(3-5 小时)增强这些基因的转录。我们的结果表明,CrhR 调节这些热休克蛋白的低温诱导表达,这对于集胞藻细胞适应低温可能是必不可少的。