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聚球藻PCC 6803中编码铁氧化还原蛋白-NADP +氧化还原酶的petH基因的表征及转录调控:一种新型反向操纵子的参与

Characterization and transcriptional regulation of the Synechocystis PCC 6803 petH gene, encoding ferredoxin-NADP+ oxidoreductase: involvement of a novel type of divergent operator.

作者信息

van Thor J J, Hellingwerf K J, Matthijs H C

机构信息

Department of Microbiology, E.C. Slater Institute, BioCentrum Amsterdam, The Netherlands.

出版信息

Plant Mol Biol. 1998 Feb;36(3):353-63. doi: 10.1023/a:1005935819038.

Abstract

The petH gene, encoding ferredoxin-NADP+ oxidoreductase (FNR), has been characterised in the unicellular cyanobacterium Synechocystis PCC 6803. Its product, FNR, was heterologously produced and functionally characterized. The start-site of the monocystronic petH transcript was mapped 523 bp upstream of the predicted PetH initiation codon, resulting in an unusually large 5'-untranslated region. The 5' end of the petH transcript is situated within the open reading frame of phosphoribulokinase (encoded by prk), which is transcribed in opposite orientation with respect to petH. The transcription start site of the prk transcript was mapped 219 bp upstream of the initiation codon, resulting in a 223 bp antisense region between both transcripts. Under many conditions the expression of both genes (i.e. petH and prk) is co-regulated symmetrically at the transcriptional level, as was concluded from both northern hybridization experiments and from primer extension analyses; it became uncoupled, however, when specifically petH expression was stimulated, independent of prk expression, by stressing the Synechocystis cells with high salt concentrations. A model for a new type of bidirectional operator, regulating the expression of petH and prk, is proposed.

摘要

编码铁氧化还原蛋白-NADP⁺氧化还原酶(FNR)的petH基因已在单细胞蓝藻集胞藻PCC 6803中得到表征。其产物FNR已通过异源表达进行了功能表征。单顺反子petH转录本的起始位点被定位在预测的PetH起始密码子上游523 bp处,导致出现一个异常大的5'非翻译区。petH转录本的5'端位于磷酸核酮糖激酶(由prk编码)的开放阅读框内,该激酶的转录方向与petH相反。prk转录本的转录起始位点被定位在起始密码子上游219 bp处,导致两个转录本之间存在一个223 bp的反义区域。从Northern杂交实验和引物延伸分析得出,在许多条件下,这两个基因(即petH和prk)的表达在转录水平上受到对称的共同调控;然而,当用高盐浓度胁迫集胞藻细胞从而特异性地刺激petH表达而不依赖于prk表达时,二者的表达变得不再偶联。本文提出了一种新型双向操纵子调控petH和prk表达的模型。

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