Levy H., Kindle K. L., Stern D. B.
Boyce Thompson Institute, Cornell University, Tower Road, Ithaca, New York 14853.
Plant Cell. 1997 May;9(5):825-836. doi: 10.1105/tpc.9.5.825.
We previously created and analyzed a Chlamydomonas reinhardtii strain, [delta]26, in which an inverted repeat in the 3[prime] untranslated region of the chloroplast atpB gene was deleted. In this strain, atpB transcripts are unstable and heterogeneous in size, and growth is poor under conditions in which photosynthesis is required. Spontaneous suppressor mutations that allow rapid photosynthetic growth have been identified. One strain, [delta]26S, retains the atpB deletion yet accumulates a discrete and stable atpB transcript as a consequence of a recessive nuclear mutation. Unlike previously isolated Chlamydomonas nuclear mutations that affect chloroplast mRNA accumulation, the mutation in [delta]26S affects several chloroplast transcripts. For example, in the atpA gene cluster, the relative abundance of several messages was altered in a manner consistent with inefficient mRNA 3[prime] end processing. Furthermore, [delta]26S cells accumulated novel transcripts with 3[prime] termini in the petD-trnR intergenic region. These transcripts are potential intermediates in 3[prime] end processing. In contrast, no alterations were detected for petD, atpA, or atpB mRNA 5[prime] ends; neither were there gross alterations detected for several other mRNAs, including the wild-type atpB transcript. We suggest that the gene identified by the suppressor mutation encodes a product involved in the processing of monocistronic and polycistronic messages.
我们之前构建并分析了莱茵衣藻(Chlamydomonas reinhardtii)菌株Δ26,该菌株中叶绿体atpB基因3′非翻译区的一个反向重复序列被删除。在这个菌株中,atpB转录本不稳定且大小不均一,并且在需要光合作用的条件下生长较差。已经鉴定出了允许快速光合生长的自发抑制突变。一个菌株Δ26S保留了atpB缺失,但由于一个隐性核突变而积累了一个离散且稳定的atpB转录本。与之前分离的影响叶绿体mRNA积累的莱茵衣藻核突变不同,Δ26S中的突变影响了几个叶绿体转录本。例如,在atpA基因簇中,几个信使RNA的相对丰度以与mRNA 3′末端加工效率低下一致的方式发生了改变。此外,Δ26S细胞在petD-trnR基因间区域积累了具有3′末端的新转录本。这些转录本是3′末端加工的潜在中间体。相比之下,未检测到petD、atpA或atpB mRNA 5′末端的改变;对于其他几个mRNA,包括野生型atpB转录本,也未检测到明显改变。我们认为,由抑制突变鉴定出的基因编码一种参与单顺反子和多顺反子信使加工的产物。