Gäbler L, Herz U, Liddell A, Leaver C J, Schröder W, Brennicke A, Grohmann L
Institut für Genbiologische Forschung Berlin, Germany.
Mol Gen Genet. 1994 Jul 8;244(1):33-40. doi: 10.1007/BF00280184.
The N-terminal amino acid sequence of a 42.5 kDa subunit of the NADH: ubiquinone oxidoreductase (complex I) from potato has been determined by direct protein sequencing. The sequence was found to be homologous to that of the nuclear-encoded 49 kDa complex I subunit of bovine and Neurospora mitochondria and to the sequence deduced from the mitochondrial nad7 gene identified in the mitochondrial (mt) DNA of tryp anosomes and the moss Marchantia. An oligonucleotide probe derived from the potato N-terminal protein sequence hybridized only to the plant mtDNA. Immunoprecipitation of in-organello 35S-labelled potato and wheat mitochondrial translation products with an antibody directed against the Neurospora 49 kDa complex I subunit indicates that at least in these plants the NAD7 protein is synthesized within the organelle. Comparisons of genomic, cDNA and protein sequences of the 5' coding region reveal three codons that are changed by RNA-editing and confirm translation of the edited transcripts in plant mitochondria. The NAD7 protein appears to undergo post-translational processing since the N-terminal methionine residue is absent from the mature mitochondrial protein.
通过直接蛋白质测序,已确定了马铃薯中NADH:泛醌氧化还原酶(复合体I)42.5 kDa亚基的N端氨基酸序列。发现该序列与牛和粗糙脉孢菌线粒体中核编码的49 kDa复合体I亚基的序列同源,也与从锥虫和地钱线粒体(mt)DNA中鉴定出的线粒体nad7基因推导的序列同源。源自马铃薯N端蛋白质序列的寡核苷酸探针仅与植物mtDNA杂交。用针对粗糙脉孢菌49 kDa复合体I亚基的抗体对离体35S标记的马铃薯和小麦线粒体翻译产物进行免疫沉淀,表明至少在这些植物中,NAD7蛋白是在细胞器内合成的。对5'编码区的基因组、cDNA和蛋白质序列进行比较,发现有三个密码子通过RNA编辑发生了改变,并证实了植物线粒体中编辑转录本的翻译。NAD7蛋白似乎经历了翻译后加工,因为成熟的线粒体蛋白中不存在N端甲硫氨酸残基。