Steinmetz Eric J, Brow David A
Department of Biomolecular Chemistry, University of Wisconsin Medical School, 1300 University Avenue, Madison, WI 53706-1532, USA.
Mol Cell Biol. 2003 Sep;23(18):6339-49. doi: 10.1128/MCB.23.18.6339-6349.2003.
Termination of transcription by RNA polymerase II (Pol II) is a poorly understood yet essential step in eukaryotic gene expression. Termination of pre-mRNA synthesis is coupled to recognition of RNA signals that direct cleavage and polyadenylation of the nascent transcript. Termination of nonpolyadenylated transcripts made by Pol II in the yeast Saccharomyces cerevisiae, including the small nuclear and small nucleolar RNAs, requires distinct RNA elements recognized by the Nrd1 protein and other factors. We have used genetic selection to characterize the terminator of the SNR13 snoRNA gene, revealing a bipartite structure consisting of an upstream element closely matching a Nrd1-binding sequence and a downstream element similar to a cleavage/polyadenylation signal. Genome-wide selection for factors influencing recogniton of the SNR13 terminator yielded mutations in the gene coding for the essential Pol II-binding protein Ssu72. Ssu72 has recently been found to associate with the pre-mRNA cleavage/polyadenylation machinery, and we find that an ssu72 mutation that disrupts Nrd1-dependent termination also results in deficient poly(A)-dependent termination. These findings extend the parallels between the two termination pathways and suggest that they share a common mechanism to signal Pol II termination.
RNA聚合酶II(Pol II)介导的转录终止是真核基因表达中一个尚未被充分理解但至关重要的步骤。前体mRNA合成的终止与对RNA信号的识别相关联,这些信号指导新生转录本的切割和聚腺苷酸化。酿酒酵母中由Pol II产生的非聚腺苷酸化转录本(包括小核RNA和小核仁RNA)的终止需要由Nrd1蛋白和其他因子识别的独特RNA元件。我们利用遗传筛选来表征SNR13小核仁RNA基因的终止子,揭示了一种二分结构,该结构由一个与Nrd1结合序列紧密匹配的上游元件和一个类似于切割/聚腺苷酸化信号的下游元件组成。全基因组筛选影响SNR13终止子识别的因子,在编码必需的Pol II结合蛋白Ssu72的基因中产生了突变。最近发现Ssu72与前体mRNA切割/聚腺苷酸化机制相关联,并且我们发现破坏Nrd1依赖性终止的ssu72突变也导致聚(A)依赖性终止缺陷。这些发现扩展了两种终止途径之间的相似性,并表明它们共享一种向Pol II发出终止信号的共同机制。