监测mRNA脱帽活性。

Monitoring mRNA decapping activity.

作者信息

Zhang S, Williams C J, Wormington M, Stevens A, Peltz S W

机构信息

Department of Molecular Genetics and Microbiology, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.

出版信息

Methods. 1999 Jan;17(1):46-51. doi: 10.1006/meth.1998.0706.

DOI:10.1006/meth.1998.0706

PMID:10075882

Abstract

mRNA decapping is a common step shared between two important mRNA decay pathways in yeast, Saccharomyces cerevisiae. To investigate how mRNAs are decapped, we have developed an assay that can be easily used to measure the decapping activity. This assay has been used to isolate yeast strains with altered decapping activities. The results demonstrated that decreased decapping activity in vitro corresponds well with the decapping-deficient phenotype in vivo. This assay has been applied to the purified yeast decapping enzyme Dcp1 protein as well as crude yeast extracts and Xenopus oocyte extracts.

摘要

mRNA去帽是酿酒酵母（Saccharomyces cerevisiae）中两条重要的mRNA降解途径共有的一个常见步骤。为了研究mRNA是如何去帽的，我们开发了一种易于用于测量去帽活性的检测方法。该检测方法已被用于分离具有改变的去帽活性的酵母菌株。结果表明，体外去帽活性的降低与体内去帽缺陷表型高度吻合。该检测方法已应用于纯化的酵母去帽酶Dcp1蛋白以及粗制酵母提取物和非洲爪蟾卵母细胞提取物。