Del Campo Eva M, Sabater Bartolomé, Martín Mercedes
Department of Plant Biology, University of Alcalá, Alcalá de Henares, 28871 Madrid, Spain.
J Biol Chem. 2002 Sep 27;277(39):36457-64. doi: 10.1074/jbc.M204500200. Epub 2002 Jul 12.
Intergenic cleavages, intron splicing, and editing of primary transcripts of the plastid ndhH-D operon produce multiple overlapping RNAs, of which the most abundant by far is the monocistronic 400-nucleotide mRNA of psaC (encoding the PsaC protein of photosystem I), in contrast with the low level of transcripts of the six ndh genes. Like other plastid operons containing genes for functionally unrelated proteins, the contrasting accumulation of ndh and psaC transcripts provides a model to investigate the mechanisms of the post-transcriptional control of gene expression, a feature of chloroplast genetic machinery, with a minimum of interference by transcriptional control. In leek (Allium porrum L), the ndhD transcript (which follows the psaC gene and ends the ndhH-D operon) requires C --> U editing to restore its start codon and may be used as a marker for the processing of psaC and ndhD transcripts. By determining the editing state and 5' end sequences of specific transcripts, we demonstrated that stable monocistronic psaC mRNA results from downstream cleavages in the ndhD sequence, which renders non-functional ndhD transcripts as by-products. Alternative psaC-ndhD intergenic cleavages produce complete mRNAs for both genes, but only take place in precursors containing editing-restored ndhD start codons. Hence, post-transcriptional control acts by promoting the ndhD cleavage alternative, which allows the accumulation of psaC mRNA at the expense of ndhD mRNA levels.
质体ndhH-D操纵子初级转录本的基因间切割、内含子剪接和编辑产生多种重叠RNA,其中迄今为止最丰富的是psaC的单顺反子400个核苷酸的mRNA(编码光系统I的PsaC蛋白),这与六个ndh基因的转录本水平较低形成对比。与其他包含功能不相关蛋白质基因的质体操纵子一样,ndh和psaC转录本的不同积累为研究基因表达的转录后控制机制提供了一个模型,这是叶绿体遗传机制的一个特征,且转录控制的干扰最小。在韭菜(葱属植物)中,ndhD转录本(位于psaC基因之后并终止ndhH-D操纵子)需要C→U编辑来恢复其起始密码子,可作为psaC和ndhD转录本加工的标记。通过确定特定转录本的编辑状态和5'端序列,我们证明稳定的单顺反子psaC mRNA是由ndhD序列中的下游切割产生的,这使得无功能的ndhD转录本成为副产物。psaC-ndhD基因间的选择性切割产生两个基因的完整mRNA,但只发生在含有编辑恢复的ndhD起始密码子的前体中。因此,转录后控制通过促进ndhD切割替代作用来发挥作用,这使得psaC mRNA得以积累,而ndhD mRNA水平降低。