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酵母scERV1基因的一个突变体在线粒体形态和分布方面表现出一种新的缺陷。

A mutant for the yeast scERV1 gene displays a new defect in mitochondrial morphology and distribution.

作者信息

Becher D, Kricke J, Stein G, Lisowsky T

机构信息

Institut für Mikrobiologie/Genetik, Ernst-Moritz-Arndt-Universität Greifswald, Jahnstrasse 15a, 17487 Greifswald, Germany.

出版信息

Yeast. 1999 Sep 15;15(12):1171-81. doi: 10.1002/(SICI)1097-0061(19990915)15:12<1171::AID-YEA443>3.0.CO;2-T.

Abstract

The yeast scERV1 gene is the best characterized representative of a new gene family found in different lower and higher eukaryotes. The gene product is essential for the yeast cell and has a complex influence on different aspects of mitochondrial biogenesis. The homologous mammalian ALR(Augmenter of Liver Regeneration) genes from man, mouse and rat are important at different developmental stages of the organism as, for example, in spermatogenesis and liver regeneration. In this study the influence of scERV1 on the morphology of mitochondria and its submitochondrial localization are investigated. A temperature-sensitive mutant of the gene was stained with a mitochondria-specific dye and fluorescence was inspected at the permissive and restrictive temperature. A new phenotype for morphological defects of mitochondria was identified. Already at the permissive temperature mitochondrial vesicles accumulate at defined positions in the cell. After shift to the restrictive temperature, morphological changes, and finally complete loss of mitochondrial structures, are observed. Ultrastructural studies confirm these findings and demonstrate the loss of the mitochondrial inner membrane and at the final stage a drastic reduction or complete absence of mitochondria from the cell. GFP fusion experiments with the scERV1 gene and subcellular localization by fractionation experiments identify the gene product inside mitoplasts and the cytosol. Re-investigation of the mutant phenotype demonstrates that after longer incubation of the mutant at the restrictive temperature an irreversible defect of the cells, even on glucose complete medium, is found that is in accordance with a complete loss or irreversible damage of mitochondria.

摘要

酵母scERV1基因是在不同的低等和高等真核生物中发现的一个新基因家族中最具特征的代表。该基因产物对酵母细胞至关重要,并且对线粒体生物发生的不同方面具有复杂的影响。来自人、小鼠和大鼠的同源哺乳动物ALR(肝脏再生增强因子)基因在生物体的不同发育阶段很重要,例如在精子发生和肝脏再生过程中。在本研究中,研究了scERV1对线粒体形态及其亚线粒体定位的影响。用线粒体特异性染料对该基因的温度敏感突变体进行染色,并在允许温度和限制温度下检查荧光。鉴定出线粒体形态缺陷的一种新表型。在允许温度下,线粒体囊泡就会在细胞内的特定位置积累。转移到限制温度后,会观察到形态变化,最终线粒体结构完全丧失。超微结构研究证实了这些发现,并证明了线粒体内膜的丧失,在最后阶段,细胞中线粒体急剧减少或完全缺失。通过scERV1基因的绿色荧光蛋白融合实验和分级分离实验进行亚细胞定位,确定了线粒体质和细胞质中的基因产物。对突变体表型的重新研究表明,在突变体于限制温度下孵育更长时间后,即使在葡萄糖完全培养基上,细胞也会出现不可逆的缺陷,这与线粒体的完全丧失或不可逆损伤一致。

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