Butler J S, Platt T
Department of Biochemistry, University of Rochester Medical Center, NY 14642.
Science. 1988 Dec 2;242(4883):1270-4. doi: 10.1126/science.2848317.
In whole cell extracts of Saccharomyces cerevisiae, incubation of precursor mRNA transcripts encoding the sequences essential in vivo for forming the 3' end of the iso-1-cytochrome c mRNA (CYC1) revealed an endonuclease activity with the characteristics required for producing the mature mRNA 3' end. The observed cleavage in vitro is (i) accurate, occurring at or near the polyadenylation site of CYC1 RNA, (ii) 30 to 50 percent efficient, (iii) adenosine triphosphate dependent, (iv) specific for the 3' ends of at least two yeast pre-mRNA's, and (v) absent with related pre-mRNA's carrying mutations that abolish correct 3' end formation in vivo. In addition, a second activity in the extract polyadenylates the product under appropriate conditions. Thus, the mature 3' ends of yeast mRNA's may be generated by endonucleolytic cleavage and polyadenylation rather than by transcription termination.
在酿酒酵母的全细胞提取物中,对编码体内形成异 - 1 - 细胞色素c mRNA(CYC1)3'端所必需序列的前体mRNA转录本进行孵育,发现了一种具有产生成熟mRNA 3'端所需特性的核酸内切酶活性。体外观察到的切割具有以下特点:(i)准确,发生在CYC1 RNA的聚腺苷酸化位点或其附近;(ii)效率为30%至50%;(iii)依赖三磷酸腺苷;(iv)对至少两种酵母前体mRNA的3'端具有特异性;(v)对于在体内消除正确3'端形成的相关前体mRNA则不存在这种活性。此外,提取物中的第二种活性在适当条件下会使产物聚腺苷酸化。因此,酵母mRNA的成熟3'端可能是通过核酸内切酶切割和聚腺苷酸化产生的,而不是通过转录终止产生的。