Isolation and characterization of pre-mRNA splicing mutants of Saccharomyces cerevisiae.
Isolation and characterization of pre-mRNA splicing mutants of Saccharomyces cerevisiae.
复制标题
酿酒酵母前体 mRNA 剪接突变体的分离和表征。
DOI：
10.1101/gad.3.8.1206
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发表时间：
1989
影响因子：
10.5
通讯作者：
Abelson,J
中科院分区：
文献类型：
--
作者：
Vijayraghavan,U;Company,M;Abelson,J
在这项研究中，我们报告的分离温度敏感的突变体，影响前mRNA剪接。一个银行的约1000个温度敏感的酿酒酵母菌株产生和筛选RNA凝胶印迹杂交与肌动蛋白内含子探针。我们分离了16个突变体，定义了11个新的互补群prp（rna）17-prp（rna）27，其中有4种表型突变体和21个prp 2-prp 11互补群（以前称为rna 2-rna 11）的突变体。大多数互补组共享前mRNA积累的表型，在所有prp（rna）2-prp（rna）11突变体中可见。在这项研究中分离出三种新的突变体。一类，由两个互补组，表现出积累的剪接的中间体，而前mRNA的水平没有变化。第二类，也由两个互补组表示，显示剪接后释放的内含子的积累。第三个新的类，包括一个互补组，积累前mRNA和释放的内含子。所有分离的突变体都是隐性的剪接表型。11个互补群中只有2个是隐性的，但对温度不敏感。这项研究，连同先前分离的prp（rna）2-prp（rna）11组和剪接体snRNAs，提出至少26个基因产物直接或间接参与前mRNA剪接。
In this study we report the isolation of temperature-sensitive mutants that affect pre-mRNA splicing. A bank of approximately 1000 temperature-sensitive Saccharomyces cerevisiae strains was generated and screened on RNA gel blots by hybridization with an actin intron probe. We isolated 16 mutants defining 11 new complementation groups prp(rna)17-prp(rna)27 with four phenotypic classes of mutants and 21 mutants in the prp2-prp11 complementation groups (formerly rna2-rna11). The majority of the complementation groups share a phenotype of pre-mRNA accumulation, seen in all of the prp(rna)2-prp(rna)11 mutants. Three novel classes of mutants were isolated in this study. One class, consisting of two complementation groups, exhibits an accumulation of the lariat intermediate of splicing, with no change in the levels of pre-mRNA. The second class, also represented by two complementation groups, shows an accumulation of the intron released after splicing. The third novel class, comprising one complementation group, accumulates both pre-mRNA and the released intron. All mutants isolated were recessive for the splicing phenotype. Only 2 of the 11 complementation groups, although recessive, were not temperature sensitive. This study, together with previous isolation of the prp(rna)2-prp(rna)11 groups and the spliceosomal snRNAs, puts at least 26 gene products involved directly or indirectly in pre-mRNA splicing.