Jiao Henry S, Hicks Amanda, Simpson Clare, Stern David B
Boyce Thompson Institute for Plant Research, Cornell University, Tower Road, Ithaca, NY 14853, USA.
Curr Genet. 2004 May;45(5):311-22. doi: 10.1007/s00294-004-0487-7. Epub 2004 Feb 4.
The Chlamydomonas reinhardtii chloroplast genome possesses thousands of small dispersed repeats (SDRs), which are of unknown function. Here, we used the petA gene as a model to investigate the role of SDRs in mRNA 3' end formation. In wild-type cells, petA mRNA accumulated as a major 1.3-kb transcript, whose 3' end was mapped to the distal end of a predicted stem-loop structure. To determine whether this stem-loop was required for petA mRNA stability, a series of deletions was constructed. These deletion strains accumulated a variety of petA mRNAs, for which approximate 3' ends were deduced. These 3' ends were found to flank stem-loop structures, many of which were formed partially or completely from inverted copies of SDRs. All strains accumulated wild-type levels of cytochrome f, demonstrating that alternative 3' termini are compatible with efficient translation. The ability to form alternative mRNA termini using SDRs lends additional flexibility to the chloroplast gene expression apparatus and thus could confer an evolutionary advantage.
莱茵衣藻叶绿体基因组拥有数千个小的分散重复序列(SDR),其功能未知。在这里,我们以petA基因为模型来研究SDR在mRNA 3'末端形成中的作用。在野生型细胞中,petA mRNA以主要的1.3 kb转录本形式积累,其3'末端定位于预测的茎环结构的远端。为了确定这个茎环是否是petA mRNA稳定性所必需的,构建了一系列缺失突变体。这些缺失突变体积累了多种petA mRNA,并推断出其大致的3'末端。发现这些3'末端位于茎环结构两侧,其中许多茎环结构部分或完全由SDR的反向拷贝形成。所有突变体中细胞色素f的水平都与野生型相当,这表明替代的3'末端与高效翻译兼容。利用SDR形成替代mRNA末端的能力为叶绿体基因表达机制提供了额外的灵活性,因此可能赋予进化优势。