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莱茵衣藻叶绿体中的petD基因由功能冗余的启动子转录。

The petD gene is transcribed by functionally redundant promoters in Chlamydomonas reinhardtii chloroplasts.

作者信息

Sturm N R, Kuras R, Büschlen S, Sakamoto W, Kindle K L, Stern D B, Wollman F A

机构信息

Boyce Thompson Institute for Plant Research at Cornell University, Ithaca, New York 14853.

出版信息

Mol Cell Biol. 1994 Sep;14(9):6171-9. doi: 10.1128/mcb.14.9.6171-6179.1994.

Abstract

FUD6, a nonphotosynthetic mutant of Chlamydomonas reinhardtii, was previously found to be deficient in the synthesis of subunit IV of the cytochrome b6/f complex, the chloroplast petD gene product (C. Lemaire, J. Girard-Bascou, F.-A. Wollman, and P. Bennoun, Biochim. Biophys. Acta 851:229-238, 1986). The lesion in FUD6 is a 236-bp deletion between two 11-bp direct repeats in the chloroplast genome. It extends from 82 to 72 bp upstream of the 5' end of wild-type petD mRNA to 156 to 166 bp downstream of the 5' end. Thus, the deletion extends into the putative promoter and 5' untranslated region of petD. No petD mRNA of the normal size can be detected in FUD6 cells, but a low level of a dicistronic message accumulates, which contains the coding regions for subunit IV and cytochrome f, the product of the upstream petA gene. petD transcriptional activity in FUD6 is not significantly altered from the wild-type level. This transcriptional activity was eliminated by petA promoter disruptions, suggesting that it originates at the petA promoter. We conclude that the petD-coding portion of most cotranscripts is rapidly degraded in FUD6, possibly following processing events that generate the 3' end of petA mRNA. A chloroplast transformant was constructed in which only the sequence from -81 to -2 relative to the major 5' end of the petD transcript was deleted. Although this deletion eliminates all detectable petD promoter activity, the transformant grows phototrophically and accumulates high levels of monocistronic petD mRNA. We conclude that the petD gene can be transcribed by functionally redundant promoters. In the absence of a functional petD promoter, a lack of transcription termination allows the downstream petD gene to be cotranscribed with the petA coding region and thereby expressed efficiently.

摘要

莱茵衣藻的非光合突变体FUD6先前被发现缺乏细胞色素b6/f复合体亚基IV(叶绿体petD基因产物)的合成能力(C. 勒梅尔、J. 吉拉尔 - 巴斯库、F.-A. 沃尔曼和P. 本努恩,《生物化学与生物物理学报》851:229 - 238,1986年)。FUD6中的损伤是叶绿体基因组中两个11 bp直接重复序列之间236 bp的缺失。它从野生型petD mRNA 5'端上游82至72 bp延伸到5'端下游156至166 bp。因此,该缺失延伸到petD的推定启动子和5'非翻译区。在FUD6细胞中检测不到正常大小的petD mRNA,但积累了低水平的双顺反子信息,其包含亚基IV和细胞色素f(上游petA基因的产物)的编码区。FUD6中petD的转录活性与野生型水平相比没有显著改变。petA启动子破坏消除了这种转录活性,表明它起源于petA启动子。我们得出结论,FUD6中大多数共转录本的petD编码部分可能在产生petA mRNA 3'端的加工事件之后迅速降解。构建了一个叶绿体转化体,其中相对于petD转录本主要5'端仅缺失了从 - 81至 - 2的序列。尽管此缺失消除了所有可检测到的petD启动子活性,但该转化体能够进行光合生长并积累高水平的单顺反子petD mRNA。我们得出结论,petD基因可以由功能冗余的启动子转录。在缺乏功能性petD启动子的情况下,转录终止的缺失使得下游的petD基因能够与petA编码区共转录,从而有效表达。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9409/359144/f26ae2d4a697/molcellb00009-0570-a.jpg

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