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首页> 外文OA文献 >Alternative Splicing Gives Rise to Different Isoforms of the Neurospora crassa Tob55 Protein That Vary in Their Ability to Insert β-Barrel Proteins Into the Outer Mitochondrial Membrane
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Alternative Splicing Gives Rise to Different Isoforms of the Neurospora crassa Tob55 Protein That Vary in Their Ability to Insert β-Barrel Proteins Into the Outer Mitochondrial Membrane

机译：选择性剪接增加了神经孢子虫Tob55蛋白的不同同工型，这些同工型将β-桶蛋白插入外部线粒体膜的能力各不相同

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摘要

Tob55 is the major component of the TOB complex, which is found in the outer membrane of mitochondria. A sheltered knockout of the tob55 gene was developed in Neurospora crassa. When grown under conditions that reduce the levels of the Tob55 protein, the strain exhibited a reduced growth rate and mitochondria isolated from these cells were deficient in their ability to import β-barrel proteins. Surprisingly, Western blots of wild-type mitochondrial proteins revealed two bands for Tob55 that differed by ∼4 kDa in their apparent molecular masses. Sequence analysis of cDNAs revealed that the tob55 mRNA is alternatively spliced and encodes three isoforms of the protein, which are predicted to contain 521, 516, or 483 amino acid residues. Mass spectrometry of proteins isolated from purified outer membrane vesicles confirmed the existence of each isoform in mitochondria. Strains that expressed each isoform of the protein individually were constructed. When cells expressing only the longest form of the protein were grown at elevated temperature, their growth rate was reduced and mitochondria isolated from these cells were deficient in their ability to assembly β-barrel proteins.
机译：Tob55是TOB复合物的主要成分，它存在于线粒体的外膜中。在neurospora crassa中开发了一个隐蔽的tob55基因敲除。当在降低Tob55蛋白水平的条件下生长时，该菌株显示出降低的生长速率，并且从这些细胞中分离出的线粒体缺乏导入β-桶形蛋白的能力。出乎意料的是，野生型线粒体蛋白的蛋白质印迹显示出Tob55的两条带，其表观分子质量相差约4 kDa。 cDNA的序列分析显示，tob55 mRNA被交替剪接并编码该蛋白质的三种同工型，预计它们包含521、516或483个氨基酸残基。从纯化的外膜囊泡分离的蛋白质的质谱分析证实了线粒体中每种同工型的存在。构建分别表达蛋白质的每种同工型的菌株。当仅表达最长形式蛋白质的细胞在高温下生长时，它们的生长速率降低，并且从这些细胞中分离出的线粒体缺乏组装β-桶形蛋白质的能力。

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