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Severe growth defect in a Schizosaccharomyces pombe mutant defective in intron lariat degradation.

机译:粟酒裂殖酵母突变体中严重的生长缺陷,内含子套索降解中存在缺陷。

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The cDNAs and genes encoding the intron lariat-debranching enzyme were isolated from the nematode Caenorhabditis elegans and the fission yeast Schizosaccharomyces pombe based on their homology with the Saccharomyces cerevisiae gene. The cDNAs were shown to be functional in an interspecific complementation experiment; they can complement an S. cerevisiae dbr1 null mutant. About 2.5% of budding yeast S. cerevisiae genes have introns, and the accumulation of excised introns in a dbr1 null mutant has little effect on cell growth. In contrast, many S. pombe genes contain introns, and often multiple introns per gene, so that S. pombe is estimated to contain approximately 40 times as many introns as S. cerevisiae. The S. pombe dbr1 gene was disrupted and shown to be nonessential. Like the S. cerevisiae mutant, the S. pombe null mutant accumulated introns to high levels, indicating that intron lariat debranching represents a rate-limiting step in intron degradation in both species. Unlike the S. cerevisiae mutant, the S. pombe dbr1::leu1+ mutant had a severe growth defect and exhibited an aberrant elongated cell shape in addition to an intron accumulation phenotype. The growth defect of the S. pombe dbr1::leu1+ strain suggests that debranching activity is critical for efficient intron RNA degradation and that blocking this pathway interferes with cell growth.
机译:根据与酿酒酵母基因的同源性,从线虫秀丽隐杆线虫和裂变酵母裂殖酵母中分离出编码内含子套索脱支酶的cDNA和基因。这些cDNA在种间互补实验中显示出功能。它们可以补充酿酒酵母dbr1 null突变体。约2.5%的酵母酿酒酵母基因具有内含子,而被切除的内含子在dbr1 null突变体中的积累对细胞生长几乎没有影响。相反,许多粟酒裂殖酵母基因含有内含子,每个基因通常含有多个内含子,因此估计粟酒裂殖酵母的内含子约为酿酒酵母的40倍。粟酒裂殖酵母dbr1基因被破坏,显示为非必需。像酿酒酵母突变体一样,粟酒裂殖酵母无效突变体将内含子积累到高水平,这表明内含子套索状脱支代表了这两个物种内含子降解的限速步骤。与酿酒酵母突变体不同,粟酒裂殖酵母dbr1 :: leu1 +突变体具有严重的生长缺陷,除内含子积累表型外,还表现出异常的细长细胞形状。粟酒裂殖酵母dbr1 :: leu1 +菌株的生长缺陷表明脱支活性对于有效的内含子RNA降解至关重要,并且阻断该途径会干扰细胞生长。

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