酵母线粒体降解体解旋酶、ATP酶和核糖核酸外切酶活性的测定。
Assays of the helicase, ATPase, and exoribonuclease activities of the yeast mitochondrial degradosome.
作者信息
Malecki Michal, Stepien Piotr P, Golik Pawel
机构信息
Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, Pawinskiego 5A, 02-106, Warsaw, Poland.
出版信息
Methods Mol Biol. 2010;587:339-58. doi: 10.1007/978-1-60327-355-8_24.
The mitochondrial degradosome (mtEXO) is the main enzymatic complex in RNA degradation, processing, and surveillance in Saccharomyces cerevisiae mitochondria. It consists of two nuclear-encoded subunits: the ATP-dependent RNA helicase Suv3p and the 3' to 5' exoribonuclease Dss1p. The two subunits depend on each other for their activity; the complex can therefore be considered as a model system for the cooperation of RNA helicases and exoribonucleases in RNA degradation. All the three activities of the complex (helicase, ATPase, and exoribonuclease) can be studied in vitro using recombinant proteins and protocols presented in this chapter.
线粒体降解体（mtEXO）是酿酒酵母线粒体中RNA降解、加工和监测的主要酶复合物。它由两个核编码亚基组成：ATP依赖性RNA解旋酶Suv3p和3'至5'外切核糖核酸酶Dss1p。这两个亚基的活性相互依赖；因此，该复合物可被视为RNA解旋酶和外切核糖核酸酶在RNA降解中协同作用的模型系统。利用本章介绍的重组蛋白和实验方案，可以在体外研究该复合物的所有三种活性（解旋酶、ATP酶和外切核糖核酸酶）。
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