Memon A R, Meng B, Mullet J E
Department of Biochemistry and Biophysics, Texas A & M University, College Station 77843, USA.
Plant Mol Biol. 1996 Mar;30(6):1195-205. doi: 10.1007/BF00019552.
The stability of the psbA mRNA increases during barley chloroplast development eventually reaching a half-life of over 40 h. Translation of psbA mRNA is also regulated in a complex way. Sequence-specific RNA binding proteins may modulate the translation or stability of the psbA mRNA during chloroplast development. UV cross-linking assays revealed that chloroplast proteins of 37 and 38 kDA bind specifically to the 3' end of psbA transcripts and not to the 5' end of psbA or rbcL transcripts. The two RNA-binding proteins were partially purified by ammonium sulfate precipitation followed by heparin agarose chromatography. Deletion and site-directed mutation analysis demonstrated that the 37/38RNPs bind in a 30 nucleotide region immediately downstream from the translation termination codon and upstream of sequences capable of forming a stem-loop structure in the 3' end of psbA transcripts. Single-base changes that diminish the binding of the 37RNP also reduce binding of the 38RNP suggesting that these proteins may bind as a heterodimer. The 37/38RNPs that bind within the 3' end of psbA transcripts could modulate transcription termination, translation or mRNA stability.
在大麦叶绿体发育过程中,psbA mRNA的稳定性增加,最终半衰期超过40小时。psbA mRNA的翻译也受到复杂的调控。序列特异性RNA结合蛋白可能在叶绿体发育过程中调节psbA mRNA的翻译或稳定性。紫外线交联试验表明,37 kDa和38 kDa的叶绿体蛋白特异性结合到psbA转录本的3'末端,而不结合到psbA或rbcL转录本的5'末端。这两种RNA结合蛋白通过硫酸铵沉淀,然后肝素琼脂糖层析进行部分纯化。缺失和定点突变分析表明,37/38RNP结合在紧接psbA转录本翻译终止密码子下游30个核苷酸区域以及psbA转录本3'末端能够形成茎环结构的序列上游。减少37RNP结合的单碱基变化也会降低38RNP的结合,这表明这些蛋白质可能以异二聚体形式结合。结合在psbA转录本3'末端的37/38RNP可能调节转录终止、翻译或mRNA稳定性。