Zeltz P, Hess W R, Neckermann K, Börner T, Kössel H
Institut für Biologie III der Universität Freiburg, Germany.
EMBO J. 1993 Nov;12(11):4291-6. doi: 10.1002/j.1460-2075.1993.tb06113.x.
Sequence analysis of amplified cDNAs derived from the maize chloroplast rpoB transcript which encodes the beta subunit of a chloroplast specific, DNA dependent RNA polymerase reveals four C-to-U editing sites clustered within 150 nucleotides of the 5' terminal region of the rpoB message. These newly identified editing sites confirm the bias of chloroplast editing for certain codon transitions and for second codon positions which both appear suggestive for an involvement of the translational apparatus in the editing process. This supposition prompted us to investigate editing of the rpoB transcript from ribosome deficient, and hence protein synthesis deficient, plastids of the barley mutant albostrians. In this mutant editing is, however, not impaired at any of the editing sites functional in the barley wild type rpoB transcript. This demonstrates that chloroplast editing is neither linked to nor dependent on the chloroplast translational apparatus. As a further consequence any peptide components required for chloroplast editing must be encoded in the nuclear genome. In spite of strong sequence conservation only three of the four editing sites identified in the maize rpoB transcript are functional in barley. This indicates that sequences surrounding an editing site alone are not sufficient as determinants for the editing process in chloroplasts, but that trans-acting templates carrying the editing information for each individual site may also be required.
对源自玉米叶绿体rpoB转录本的扩增cDNA进行序列分析,该转录本编码叶绿体特异性的、依赖DNA的RNA聚合酶的β亚基,结果显示在rpoB信息5'末端区域的150个核苷酸内聚集有四个C到U的编辑位点。这些新确定的编辑位点证实了叶绿体编辑对某些密码子转换以及对第二密码子位置的偏好,这两者似乎都暗示了翻译装置参与编辑过程。这一推测促使我们研究大麦突变体白条纹叶中核糖体缺陷因而蛋白质合成缺陷的质体中rpoB转录本的编辑情况。然而,在这个突变体中,大麦野生型rpoB转录本中任何一个起作用的编辑位点的编辑都没有受损。这表明叶绿体编辑既不与叶绿体翻译装置相关,也不依赖于叶绿体翻译装置。进一步的结果是,叶绿体编辑所需的任何肽成分都必须由核基因组编码。尽管序列保守性很强,但在玉米rpoB转录本中确定的四个编辑位点中只有三个在大麦中起作用。这表明仅编辑位点周围的序列不足以作为叶绿体编辑过程的决定因素,还可能需要携带每个单独位点编辑信息的反式作用模板。