Reconstitution of Targeted Deadenylation by the Ccr4-Not Complex and the YTH Domain Protein Mmi1

被引：54

作者：

Stowell, James A. W. ^{[1
]}

Webster, Michael W. ^{[1
]}

Koegel, Alexander ^{[1
]}

Wolf, Jana ^{[1
]}

Shelley, Kathryn L. ^{[1
]}

Passmore, Lori A. ^{[1
]}

机构：

[1] MRC Lab Mol Biol, Cambridge CB2 0QH, England

来源：

CELL REPORTS | 2016年 / 17卷 / 08期

基金：

英国医学研究理事会; 欧洲研究理事会;

关键词：

MESSENGER-RNA STABILITY; FISSION YEAST; STRUCTURAL BASIS; HETEROCHROMATIN FORMATION; TRANSLATIONAL REPRESSION; SACCHAROMYCES-CEREVISIAE; MEDIATED DEADENYLATION; CRYSTAL-STRUCTURE; GENE-EXPRESSION; NUCLEASE MODULE;

D O I：

10.1016/j.celrep.2016.10.066

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

Ccr4-Not is a conserved protein complex that shortens the 3' poly(A) tails of eukaryotic mRNAs to regulate transcript stability and translation into proteins. RNA-binding proteins are thought to facilitate recruitment of Ccr4-Not to certain mRNAs, but lack of an in-vitro-reconstituted system has slowed progress in understanding the mechanistic details of this specificity. Here, we generate a fully recombinant Ccr4-Not complex that removes poly(A) tails from RNA substrates. The intact complex is more active than the exonucleases alone and has an intrinsic preference for certain RNAs. The RNA-binding protein Mmi1 is highly abundant in preparations of native Ccr4-Not. We demonstrate a high-affinity interaction between recombinant Ccr4-Not and Mmi1. Using in vitro assays, we show that Mmi1 accelerates deadenylation of target RNAs. Together, our results support a model whereby both RNA-binding proteins and the sequence context of mRNAs influence deadenylation rate to regulate gene expression.

引用

页码：1978 / 1989

页数：12