Graduate School of Natural Sciences, Nagoya City University, Yamanohata, Mizuho, Nagoya 467-8501, Japan.
Nucleic Acids Res. 2011 Nov;39(21):9405-12. doi: 10.1093/nar/gkr644. Epub 2011 Aug 16.
The chloroplast atpB and atpE genes encode subunits β and ε of the ATP synthase, respectively. They are co-transcribed as dicistronic mRNAs in flowering plants. An unusual feature is an overlap (AUGA) of the atpB stop codon (UGA) with the atpE start codon (AUG). Hence, atpE translation has been believed to depend on atpB translation (i.e. translational coupling). Using an in vitro translation system from tobacco chloroplasts, we showed that both atpB and atpE cistrons are translated from the tobacco dicistronic mRNA, and that the efficiency of atpB translation is higher than that of atpE translation. When the atpB 5'-UTR was replaced with lower efficiency 5'-UTRs, atpE translation was higher than atpB translation. Removal of the entire atpB 5'-UTR arrested atpB translation but atpE translation still proceeded. Introduction of a premature stop codon in the atpB cistron did not abolish atpE translation. These results indicate that atpE translation is independent of atpB translation. Mutation analysis showed that the atpE cistron possesses its own cis-element(s) for translation, located ~25 nt upstream from the start codon.
叶绿体 atpB 和 atpE 基因分别编码 ATP 合酶的亚基 β 和 ε。它们在开花植物中作为双顺反子 mRNA 共转录。一个不寻常的特征是 atpB 终止密码子(UGA)与 atpE 起始密码子(AUG)重叠(AUGA)。因此,一直认为 atpE 翻译依赖于 atpB 翻译(即翻译偶联)。使用来自烟草叶绿体的体外翻译系统,我们表明 atpB 和 atpE 顺式基因都从烟草双顺反子 mRNA 翻译,并且 atpB 翻译的效率高于 atpE 翻译的效率。当 atpB 5'-UTR 被效率较低的 5'-UTR 取代时,atpE 翻译的效率高于 atpB 翻译的效率。去除整个 atpB 5'-UTR 会阻止 atpB 翻译,但 atpE 翻译仍会继续进行。在 atpB 顺式基因中引入过早的终止密码子不会消除 atpE 翻译。这些结果表明 atpE 翻译独立于 atpB 翻译。突变分析表明,atpE 顺式基因具有其自身的翻译顺式元件(s),位于起始密码子上游约 25nt。