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粗糙脉孢菌萌发分生孢子中线粒体基因的表达。

Expression of mitochondrial genes in the germinating conidia of Neurospora crassa.

作者信息

Bittner-Eddy P, Monroy A F, Brambl R

机构信息

Department of Plant Biology, University of Minnesota Saint Paul 55108.

出版信息

J Mol Biol. 1994 Jan 21;235(3):881-97. doi: 10.1006/jmbi.1994.1046.

Abstract

The germinating asexual spores (conidia) of Neurospora crassa were employed to study steps in the accumulation of transcripts of groups of mitochondrial genes, including those for peptide subunits of cytochrome c oxidase (CO), ATPase (ATP), and apocytochrome b (COB). Physically clustered groups of genes were expressed as cohorts: transcripts of the ATP8-ATP6-mtATP9-CO2 genes were almost undetectable in the dormant spores, and they accumulated rapidly as a group immediately after spore activation. Transcripts of COB and the adjacent CO1 were abundant in the dormant spores, and the dormant and germinating spores contained size forms of the COB transcripts that were not evident in vegetative cells. Polyribosomes were prepared from mitochondrial lysates, and the polyribosomal RNA was probed to identify the mRNAs of specific genes; in several instances polycistronic mRNAs were present in the polyribosomes as were the smaller end-products of the inferred transcript processing pathways. The expression of the physically dispersed genes for subunit peptides of cytochrome c oxidase appears to be regulated to the level of translation; these transcripts are accumulated in the total mitochondrial RNA with sharply different kinetics, but they appeared in the polyribosomes uniformly, their appearance correlating with the uniform synthesis of the subunit peptides. Transcripts for a previously reported non-functional mitochondrial gene, homologous to the functional nuclear gene for ATPase subunit 9, were found in the germinating spores, but were not detected in vegetative cells. These mtATP9 transcripts were also present in the polyribosomes and were apparently translated into a protein in vivo whose synthesis was insensitive to cycloheximide and detectable with an anti-ATP9 subunit antibody. Transcripts for two nuclear genes for mitochondrially localized proteins, ATP9 and CO5, were accumulated in unison and especially rapidly during spore germination.

摘要

粗糙脉孢菌的无性萌发孢子(分生孢子)被用于研究线粒体基因组转录本积累的步骤,这些基因组包括细胞色素c氧化酶(CO)、ATP酶(ATP)和脱辅基细胞色素b(COB)的肽亚基基因。物理上成簇的基因组作为一个群体表达:ATP8 - ATP6 - mtATP9 - CO2基因的转录本在休眠孢子中几乎检测不到,在孢子激活后它们作为一个群体迅速积累。COB和相邻的CO1的转录本在休眠孢子中丰富,并且休眠和萌发的孢子含有在营养细胞中不明显的COB转录本大小形式。从线粒体裂解物中制备多核糖体,并对多核糖体RNA进行检测以鉴定特定基因的mRNA;在几个实例中,多顺反子mRNA存在于多核糖体中,推测的转录本加工途径的较小终产物也存在。细胞色素c氧化酶亚基肽的物理分散基因的表达似乎在翻译水平受到调控;这些转录本在线粒体总RNA中以截然不同的动力学积累,但它们在多核糖体中均匀出现,其出现与亚基肽的均匀合成相关。在萌发孢子中发现了一个先前报道的无功能线粒体基因的转录本,该基因与ATP酶亚基9的功能核基因同源,但在营养细胞中未检测到。这些mtATP9转录本也存在于多核糖体中,并且显然在体内被翻译成一种蛋白质,其合成对环己酰亚胺不敏感,并且可以用抗ATP9亚基抗体检测到。两个线粒体定位蛋白的核基因ATP9和CO5的转录本在孢子萌发期间一致且特别迅速地积累。

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