Chaudhuri S, Carrer H, Maliga P
Waksman Institute, Rutgers, The State University of New Jersey, Piscataway 08855-0759, USA.
EMBO J. 1995 Jun 15;14(12):2951-7. doi: 10.1002/j.1460-2075.1995.tb07295.x.
In tobacco plastids, functional psbL mRNA is created by editing an ACG codon to an AUG translation initiation codon. To determine if editing may occur in a chimeric mRNA, the N-terminal part of psbL containing the editing site was translationally fused with the aadA and kan bacterial genes. The chimeric constructs were introduced into the tobacco plastid genome by targeted gene insertion. Editing of the chimeric mRNAs indicated that the 98 nt fragment spanning the psbL editing site contains all cis information required for editing. Expression of the chimeric gene transcripts led to a significant decrease in the editing efficiency of the endogenous psbL mRNA. However, the efficiency of editing in the transplastomic lines was unchanged for four sites in the rpoB and ndhB mRNAs. Reduced efficiency of psbL editing, but not of the other four sites, in the transplastomic lines indicates depletion of psbL-specific editing factor(s). This finding implicates the involvement of site-specific factors in editing of plastid mRNAs in higher plants.
在烟草质体中,功能性psbL mRNA是通过将ACG密码子编辑为AUG翻译起始密码子产生的。为了确定编辑是否会在嵌合mRNA中发生,将含有编辑位点的psbL的N端部分与aadA和kan细菌基因进行翻译融合。通过靶向基因插入将嵌合构建体导入烟草质体基因组。嵌合mRNA的编辑表明,跨越psbL编辑位点的98 nt片段包含编辑所需的所有顺式信息。嵌合基因转录本的表达导致内源性psbL mRNA的编辑效率显著降低。然而,转质体系中rpoB和ndhB mRNA的四个位点的编辑效率没有变化。转质体系中psbL编辑效率降低,但其他四个位点未降低,这表明psbL特异性编辑因子耗竭。这一发现暗示了位点特异性因子参与高等植物质体mRNA的编辑。