Piper P W, Bellatin J A, Lockheart A
Department of Biochemistry, University College London, UK.
EMBO J. 1983;2(3):353-9. doi: 10.1002/j.1460-2075.1983.tb01430.x.
Sequences at the immediate 3' terminus of several eukaryotic primary transcripts, synthesised just before the termination of transcription, are often lost during RNA processing. The rna82.1 mutation in Saccharomyces cerevisiae appears to result in a deficiency of the endonuclease that removes such sequences from certain yeast transcripts. Some small RNAs of rna82.1 cells are a few nucleotides longer than their counterparts in wild-type S. cerevisiae. The 5S rRNAs made during very short pulse-labellings of the mutant have, relative to the mature 121 nucleotide 5S RNA of wild-type cells, an additional 7, 11 or 13 nucleotides at their 3' terminus. These 5S forms reveal sites upon 5S genes where transcription probably terminates in vivo. The extra nucleotides upon 5S RNAs in rna82.1 cells are lost very slowly by sequential removal from the 3' terminus. Through this 3'-5' exonuclease action the total 5S RNA of the mutant possesses several 3'-terminal sequences yet is mostly only 0-3 nucleotides longer than in wild-type S. cerevisiae. Just one or two of these 3'-terminal sequences serve as a substrate in vivo for a poly(A) polymerase since a small proportion of rna82.1 5S RNAs terminate in the sequence: CAAUCUUU(A)n.
几种真核生物初级转录本紧邻3'末端的序列,在转录终止前不久合成,在RNA加工过程中常常丢失。酿酒酵母中的rna82.1突变似乎导致一种核酸内切酶缺陷,该酶负责从某些酵母转录本中去除此类序列。rna82.1细胞中的一些小RNA比野生型酿酒酵母中的对应小RNA长几个核苷酸。在对突变体进行非常短时间的脉冲标记期间产生的5S rRNA,相对于野生型细胞成熟的121个核苷酸的5S RNA,在其3'末端有额外的7、11或13个核苷酸。这些5S形式揭示了5S基因上转录在体内可能终止的位点。rna82.1细胞中5S RNA上的额外核苷酸通过从3'末端顺序去除而非常缓慢地丢失。通过这种3'-5'核酸外切酶作用,突变体的总5S RNA具有几个3'-末端序列,但大多仅比野生型酿酒酵母中的长0 - 3个核苷酸。这些3'-末端序列中只有一两个在体内作为多聚腺苷酸聚合酶的底物,因为一小部分rna82.1 5S RNA以序列:CAAUCUUU(A)n终止。