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噬菌体T7的DNA注入细胞是由一种类似酶的机制启动的。

Bacteriophage T7 DNA ejection into cells is initiated by an enzyme-like mechanism.

作者信息

Kemp Priscilla, Gupta Manisha, Molineux Ian J

机构信息

Section of Molecular Genetics and Microbiology, and Institute for Cell and Molecular Biology, University of Texas, Austin, TX 78712-1095, USA.

出版信息

Mol Microbiol. 2004 Aug;53(4):1251-65. doi: 10.1111/j.1365-2958.2004.04204.x.

Abstract

In a normal infection about 850 bp of the bacteriophage T7 genome is ejected into the cell, the remainder of the genome is internalized through transcription by Escherichia coli and then T7 RNA polymerase. Rates of T7 DNA internalization by the E. coli enzyme in vivo are constant across the whole genome. As expected for an enzyme-catalysed reaction, rates vary with temperature and can be fitted to Arrhenius kinetics. Phage virions containing a mutant gp16, a protein known to be ejected from the phage capsid into the cell at the initiation of infection, allow complete entry of the T7 genome in the absence of transcription. The kinetics of DNA ejection from such a mutant virion into the bacterial cytoplasm have also been measured at different temperatures in vivo. Between 15 and 43 degrees C the entire 40 kb T7 genome is translocated into the cell at a constant rate that is characteristic for each temperature, and the temperature-dependence of DNA translocation rates can be fitted to Arrhenius kinetics. The data are consistent with the idea that transcription-independent DNA translocation from the T7 virion is also enzyme-catalysed. The proton motive force is necessary for this mode of DNA translocation, because collapsing the membrane potential while the T7 genome is entering the cell abruptly halts further DNA transfer.

摘要

在正常感染过程中,噬菌体T7基因组约850 bp的片段被注入细胞,基因组的其余部分通过大肠杆菌转录以及随后的T7 RNA聚合酶作用内化。大肠杆菌酶在体内对T7 DNA的内化速率在整个基因组中是恒定的。正如酶催化反应所预期的那样,内化速率随温度变化,并且可以拟合为阿伦尼乌斯动力学。含有突变型gp16(一种已知在感染开始时从噬菌体衣壳中注入细胞的蛋白质)的噬菌体病毒粒子,在没有转录的情况下允许T7基因组完全进入。也已在体内不同温度下测量了这种突变病毒粒子的DNA注入细菌细胞质的动力学。在15至43摄氏度之间,整个40 kb的T7基因组以每个温度特有的恒定速率转运到细胞中,并且DNA转运速率的温度依赖性可以拟合为阿伦尼乌斯动力学。这些数据与以下观点一致,即T7病毒粒子中与转录无关的DNA转运也是由酶催化的。质子动力势对于这种DNA转运模式是必需的,因为在T7基因组进入细胞时使膜电位崩溃会突然停止进一步的DNA转移。

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