Carroll Kristina L, Ghirlando Rodolfo, Ames Jessica M, Corden Jeffry L
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
RNA. 2007 Mar;13(3):361-73. doi: 10.1261/rna.338407. Epub 2007 Feb 5.
Yeast RNA-binding proteins Nrd1 and Nab3 direct transcription termination of sn/snoRNA transcripts, some mRNA transcripts, and a class of intergenic and anti-sense transcripts. Recognition of Nrd1- and Nab3-binding sites is a critical first step in the termination and subsequent processing or degradation of these transcripts. In this article, we describe the purification and characterization of an Nrd1-Nab3 heterodimer. This Nrd1-Nab3 complex binds specifically to RNA sequences derived from a snoRNA terminator. The relative binding to mutant terminators correlates with the in vivo termination efficiency of these mutations, indicating that the primary specificity determinant in nonpoly(A) termination is Nrd1-Nab3 binding. In addition, several snoRNA terminators contain multiple Nrd1- and Nab3-binding sites and we show that multiple heterodimers bind cooperatively to one of these terminators in vitro.
酵母RNA结合蛋白Nrd1和Nab3指导sn/snoRNA转录本、一些mRNA转录本以及一类基因间和反义转录本的转录终止。识别Nrd1和Nab3结合位点是这些转录本终止以及后续加工或降解的关键第一步。在本文中,我们描述了Nrd1-Nab3异二聚体的纯化和特性。这种Nrd1-Nab3复合物特异性结合源自snoRNA终止子的RNA序列。与突变终止子的相对结合与这些突变在体内的终止效率相关,表明非聚腺苷酸化(nonpoly(A))终止中的主要特异性决定因素是Nrd1-Nab3结合。此外,几个snoRNA终止子包含多个Nrd1和Nab3结合位点,并且我们表明多个异二聚体在体外协同结合其中一个终止子。