Vijayraghavan U, Company M, Abelson J
Division of Biology, California Institute of Technology, Pasadena 91125.
Genes Dev. 1989 Aug;3(8):1206-16. doi: 10.1101/gad.3.8.1206.
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.
在本研究中，我们报告了影响前体mRNA剪接的温度敏感突变体的分离。构建了一组约1000个温度敏感型酿酒酵母菌株库，并通过与肌动蛋白内含子探针杂交在RNA凝胶印迹上进行筛选。我们分离出16个突变体，它们定义了11个新的互补群prp(rna)17 - prp(rna)27，具有四类突变体表型，以及prp2 - prp11互补群（以前的rna2 - rna11）中的21个突变体。大多数互补群具有前体mRNA积累的表型，在所有prp(rna)2 - prp(rna)11突变体中都能看到。在本研究中分离出了三类新的突变体。一类由两个互补群组成，表现出剪接套索中间体的积累，前体mRNA水平没有变化。第二类同样由两个互补群代表，显示出剪接后释放的内含子的积累。第三类新的突变体由一个互补群组成，积累前体mRNA和释放的内含子。所有分离出的突变体的剪接表型均为隐性。11个互补群中只有2个，虽然是隐性的，但不是温度敏感型。这项研究，连同之前对prp(rna)2 - prp(rna)11组和剪接体snRNA的分离，确定了至少26种直接或间接参与前体mRNA剪接的基因产物。
