Developmental expression of nuclear genes that encode mitochondrial proteins: insect cytochromes c.
Developmental expression of nuclear genes that encode mitochondrial proteins: insect cytochromes c.
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DOI：
10.1073/pnas.82.7.1964
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发表时间：
1985-04
影响因子：
11.1
通讯作者：
M. Swanson;S. M. Zieminn;D. Miller;E. Garber;E. Margoliash
中科院分区：
文献类型：
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作者：
M. Swanson;S. M. Zieminn;D. Miller;E. Garber;E. Margoliash
为了研究线粒体生物发生的组织特异性发育调控，我们研究了烟草角蛾（Manduca sexta）胸肌细胞色素c基因在成虫羽化过程中的表达，并利用这一信息获得了该基因的cDNA克隆，进而用于分离黑腹果蝇（Drosophila melanogaster）相应的基因。在成虫羽化前3天，线粒体内膜的电子密度逐渐增大，呈片层状，虽然细胞色素c没有积累，但每只昆虫胸腔的全蛋白含量增加了40倍。体外翻译和印迹杂交分析表明，胸肌细胞色素c基因主要在转录水平受到调控，细胞色素c mRNA占总poly(A)+ RNA的比例从不到0.01%上升到0.04%，并在羽化后第2天下降到检测不到的水平。此外，细胞色素c与其他细胞色素的比例始终保持不变，这表明呼吸链的这些组成部分遵循协调的发育程序。采用多聚体免疫吸附法，从胸肌组织中分离出大于或等于95%细胞色素c mRNA的poly(a)+ RNA群体，构建cDNA文库，通过杂交选择/翻译筛选。我们报道了其中一个克隆pMSc750的序列，并将其用于分离果蝇的主要胸肌细胞色素c基因。
To investigate tissue-specific developmental regulation of mitochondrial biogenesis, we studied the expression of the Manduca sexta (tobacco hornworm moth) thoracic muscle cytochrome c gene during adult eclosion and used this information to obtain a cDNA clone for this gene, which in turn was used to isolate the corresponding Drosophila melanogaster gene. Over the 3 days prior to adult Manduca emergence, mitochondrial inner membranes become progressively more electron dense and lamellar, and, while there is no accumulation of apocytochrome c, the amount of the holoprotein increases 40-fold per insect thorax. As determined by in vitro translation and blot hybridization analysis, the major thoracic muscle cytochrome c gene is primarily regulated at the transcriptional level, with cytochrome c mRNA increasing from less than 0.01% to 0.04% of total poly(A)+ RNA and declining to an undetectable level by day 2 after eclosion. Furthermore, the ratio of cytochrome c to the other cytochromes remains the same at all times, indicating that these components of the respiratory chain follow coordinated developmental programs. By using polysome immunoadsorption, a poly(A)+ RNA population of greater than or equal to 95% cytochrome c mRNA was isolated from thoracic muscle tissue and was used to construct a cDNA library, which was screened by hybrid selection/translation. We report the sequence of one of those clones, pMSc750, and its use to isolate the major thoracic muscle cytochrome c gene of Drosophila.