DnaK Heat Shock Protein of Escherichia coli Maintains the Negative Supercoiling of DNA against Thermal Stress*

DnaK Heat Shock Protein of Escherichia coli Maintains the Negative Supercoiling of DNA against Thermal Stress*
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大肠杆菌的 DnaK 热休克蛋白可维持 DNA 的负超螺旋以抵抗热应激*

DOI:
--
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发表时间:
1996
影响因子:
4.8
通讯作者:
K. Sekimizu
K. Sekimizu
中科院分区:
生物学2区
文献类型:
--
作者:
Y. Ogata;T. Mizushima;K. Kataoka;K. Kita;T. Miki;K. Sekimizu

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指数生长的大肠杆菌质粒DNA在热休克后立即松弛,在热休克条件下DNA的松弛状态迅速恢复到原始状态。我们现在已经获得了遗传和生化证据,表明大肠杆菌的DnaK热休克蛋白(hsp70的原核同源物)参与了DNA的重新超卷曲。由于rpoH琥珀突变体中没有发生DNA的重新超卷曲,因此可能需要热休克蛋白来进行该反应。DNA缺失突变体的质粒DNA在温度升高后过度松弛,没有观察到DNA的重新超卷曲。用dnaK突变体制备的粗细胞提取物孵育的DNA比用其等基因野生型菌株的提取物孵育的DNA更松弛,并且在突变体提取物中添加纯化的dnaK蛋白导致DNA的负超卷曲增加。此外,纯化DNA回转酶的反应产物在DnaK蛋白存在下更负向超旋。基于这些结果,我们提出DnaK蛋白在维持DNA对热应激的负超卷曲中起作用。
Plasmid DNA in exponentially growing Escherichia coli immediately relaxes after heat shock, and the relaxed state of DNA rapidly reverts to the original state with exposure to conditions of heat shock. We have now obtained genetic and biochemical evidence indicating that DnaK heat shock protein of E. coli, a prokaryotic homologue of hsp70, is involved in this re-supercoiling of DNA. As re-supercoiling of DNA did not occur in an rpoH amber mutant, it seems likely that heat shock proteins are required for this reaction. Plasmid DNA in a dnaK deletion mutant relaxed excessively after temperature shift-up, and the re-supercoiling of DNA was not observed. DNAs incubated with a crude cell extract prepared from the dnaK mutant were more relaxed than seen with the extract from its isogenic wild-type strain, and the addition of purified DnaK protein to the mutant extract led to an increase in the negative supercoiling of DNA. Moreover, reaction products of purified DNA gyrase more negatively supercoiled in the presence of DnaK protein. Based on these results, we propose that DnaK protein plays a role in maintaining the negative supercoiling of DNA against thermal stress.
DOI: 10.1016/s0006-291x(81)80257-4
发表时间: 1981
影响因子: 3.1
作者:
Neidhardt,FC;VanBogelen,RA
通讯作者: VanBogelen,RA
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发表时间: 1987-12
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DOI: --
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影响因子: --
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DOI: --
发表时间: 1993
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影响因子: --
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大肠杆菌中 DNA 复制起始过程中 GrpE 和 DnaK 热休克蛋白激活 DnaA5 蛋白。
DOI: --
发表时间: 1993
期刊: The Journal of biological chemistry
影响因子: --
作者:
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通讯作者: Kaguni,JM