文章摘要
粟酒裂殖酵母中ppr基因缺失引起细胞絮凝的机制研究
Induction Mechanism of ppr Gene Deletion Triggering Flocculation in Schizosaccharomyces pombe
  
DOI:doi:10.3969/j.issn.1005-7021.2019.02.002
中文关键词: 絮凝  酵母  粟酒裂殖酵母  线粒体  PPR蛋白
英文关键词: flocculation  yeast  Schizosaccharomyces pombe  mitochondria  PPR protin
基金项目:国家自然基金青年基金项目(31400032);江苏省高校自然科学研究项目(13KJB180010)
作者单位
张娟 南华大学 生药学与生物科学学院,湖南 衡阳 421001 
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中文摘要:
      粟酒裂殖酵母中PPR蛋白在线粒体和叶绿体的RNA代谢中起重要作用,ppr基因的缺失会引起线粒体功能异常(ppr5除外)。旨在揭示粟酒裂殖酵母中ppr基因缺失引起细胞絮凝的机制。利用含有半乳糖的培养基分别培养WT、Δppr3、Δppr4、Δppr6和Δppr10,观察野生型和突变体的生长表型;通过添加不同的金属离子及改变细胞表面pH来探究诱导这类突变体出现絮凝的因素;采用RT-PCR技术检测缺陷菌中编码细胞表面絮凝因子及细胞壁重构酶基因的表达情况。研究结果表明:ppr3、ppr4、ppr6和ppr10的敲除会引起由Ca2+依赖型半乳糖结合蛋白参与的细胞絮凝;不同的pH和不同金属离子对这些缺陷菌的细胞絮凝有影响;ppr3、ppr4、ppr6和ppr10的缺失会使细胞内编码絮凝因子和细胞壁重构酶基因表达上调。线粒体功能异常会诱导絮凝基因表达上调和细胞壁属性发生改变,从而产生絮凝现象。
英文摘要:
      Pentatricopeptide repeat (PPR) proteins plays important role in mitochondrial and chloroplast in Schizosaccharomyces pombe RNA metabolism, the deletion of ppr gene will cause functional anomaly of mitochondria (expect for ppr5). The aim of this paper is to reveal the mechanism of ppr gene deletion that triggering cell flocculation. Disruption of ppr gene result in mitochondrial dysfunction(except ppr5). This paper aims to reveal the mechanism of cell flocculation caused by loss of ppr gene in Schizosaccharomyces pombe. The growth phenotypes of wild type and mutant strains, wild type, Δppr3, Δppr4, Δppr6 and Δppr10 were respectively observed culturing in medium containing galactose; factors that induced the flocculation of these mutants by adding different metal ions and changing the cell surface pH were investigated; adopting RT-PCR technique to detect the expression of cell surface flocculation factor and cell wall remodeling enzyme genes in defect strains. The results of the study showed that the knocked-off of ppr3, ppr4, ppr6, and ppr10 will cause the calcium dependent galactose binding proteins to take part in cell flocculation; different pH and different metal ions had effects on cell flocculation of these defect strains; the deletion of ppr3, ppr4, ppr6, and ppr10 will make the expressions of flocculation genes and cell-wall remodeling enzyme genes up regulated. To summarize the above mentioned, the functional anomaly of mitochondria may induce flocculation expression up regulated and the changes of cell wall, thereby leading to the appearance of flocculation.
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