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RpiR 样阻遏蛋白 IolR 调控苜蓿中华根瘤菌肌醇代谢。

The RpiR-like repressor IolR regulates inositol catabolism in Sinorhizobium meliloti.

机构信息

Department of Biological Sciences, Western Michigan University, Kalamazoo, MI 49008-5410, USA.

出版信息

J Bacteriol. 2011 Oct;193(19):5155-63. doi: 10.1128/JB.05371-11. Epub 2011 Jul 22.

Abstract

Sinorhizobium meliloti, the nitrogen-fixing symbiont of alfalfa, has the ability to catabolize myo-, scyllo-, and D-chiro-inositol. Functional inositol catabolism (iol) genes are required for growth on these inositol isomers, and they play a role during plant-bacterium interactions. The inositol catabolism genes comprise the chromosomally encoded iolA (mmsA) and the iolY(smc01163)RCDEB genes, as well as the idhA gene located on the pSymB plasmid. Reverse transcriptase assays showed that the iolYRCDEB genes are transcribed as one operon. The iol genes were weakly expressed without induction, but their expression was strongly induced by myo-inositol. The putative transcriptional regulator of the iol genes, IolR, belongs to the RpiR-like repressor family. Electrophoretic mobility shift assays demonstrated that IolR recognized a conserved palindromic sequence (5'-GGAA-N6-TTCC-3') in the upstream regions of the idhA, iolY, iolR, and iolC genes. Complementation assays found IolR to be required for the repression of its own gene and for the downregulation of the idhA-encoded myo-inositol dehydrogenase activity in the presence and absence of inositol. Further expression studies indicated that the late pathway intermediate 2-keto-5-deoxy-D-gluconic acid 6-phosphate (KDGP) functions as the true inducer of the iol genes. The iolA (mmsA) gene encoding methylmalonate semialdehyde dehydrogenase was not regulated by IolR. The S. meliloti iolA (mmsA) gene product seems to be involved in more than only the inositol catabolic pathway, since it was also found to be essential for valine catabolism, supporting its more recent annotation as mmsA.

摘要

苜蓿中华根瘤菌是紫花苜蓿的固氮共生体,能够分解肌醇、鲨肌醇和 D-手性肌醇。在这些肌醇异构体上生长需要功能性肌醇分解代谢(iol)基因,这些基因在植物-细菌相互作用中发挥作用。肌醇分解代谢基因包括染色体编码的 iolA(mmsA)和 iolY(smc01163)RCDEB 基因,以及位于 pSymB 质粒上的 idhA 基因。逆转录酶分析表明,iolYRCDEB 基因作为一个操纵子转录。iol 基因在没有诱导的情况下表达较弱,但在肌醇诱导下表达强烈。iol 基因的假定转录调节剂 IolR 属于 RpiR 样阻遏物家族。电泳迁移率变动分析表明,IolR 识别 idhA、iolY、iolR 和 iolC 基因上游区域中保守的回文序列(5'-GGAA-N6-TTCC-3')。互补测定发现,IolR 是自身基因抑制和在有或没有肌醇的情况下下调 idhA 编码的肌醇脱氢酶活性所必需的。进一步的表达研究表明,晚期途径中间产物 2-酮-5-脱氧-D-葡萄糖酸 6-磷酸(KDGP)是 iol 基因的真正诱导物。编码甲基丙二酰半醛脱氢酶的 iolA(mmsA)基因不受 IolR 调控。苜蓿中华根瘤菌 iolA(mmsA)基因产物似乎不仅参与肌醇分解代谢途径,因为它还被发现对缬氨酸分解代谢至关重要,这支持了其最近被注释为 mmsA。

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