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聚球藻7942中假定的铁硫蛋白IdiC(开放阅读框5)的特性分析

Characterization of the putative iron sulfur protein IdiC (ORF5) in Synechococcus elongatus PCC 7942.

作者信息

Pietsch Daniel, Staiger Dorothee, Pistorius Elfriede K, Michel Klaus-Peter

机构信息

Lehrstuhl für Molekulare Zellphysiologie, Fakultät für Biologie, Universität Bielefeld, Universitätsstr. 25, 33615 Bielefeld, Germany.

出版信息

Photosynth Res. 2007 Oct;94(1):91-108. doi: 10.1007/s11120-007-9222-9. Epub 2007 Aug 10.

Abstract

The IdiC protein (iron deficiency induced protein C) is encoded by orf5 (now called idiC), which is part of the iron-responsive idiB operon of Synechococcus elongatus PCC 7942. The 20.5 kDa IdiC protein has a putative transmembrane helix and belongs to the thioredoxin (TRX)-like [2Fe-2S] ferredoxin family. IdiC has the highest similarity to the peripheral subunit NuoE of the Escherichia coli NDH-1 complex. IdiC expression increased under iron starvation and also in the late growth phase, representing growth conditions, which favor photosynthetic cyclic and respiratory electron transport over photosynthetic linear electron transport from water to NADP+. Attempts to insertionally inactivate the idiC gene generated merodiploid mutants with a strongly reduced IdiC content (mutant MuD) but no IdiC-free mutant. Thus, IdiC seems to be an essential protein for the viability of S. elongatus under the used experimental conditions. Comparative analyses of S. elongatus wild type (WT) and mutant MuD showed that under iron limitation in WT and MuD the amount of the reaction center proteins PsbA and PsaA/B was highly reduced. MuD had a lower growth rate, chlorophyll content, and photosynthetic O2 evolving activity with bicarbonate as electron acceptor than WT. Immunoblot analyses also showed that in MuD, when grown under iron limitation, the amount of the proteins IdiC and IdiB was greatly reduced as compared to WT. As a consequence of the reduction of the transcription factor IdiB, IdiA and IrpA expression were also decreased. In addition, the IsiA protein concentration was lower in MuD than in WT, although the isiA mRNA was equally high in MuD and WT. Another significant difference was the lower expression of the ferredoxin:NADP+ oxidoreductase in mutant MuD under iron limitation compared to WT. A possible function of the protein IdiC in cyclic electron transport around photosystem I and/or in respiratory electron transport will be discussed.

摘要

IdiC蛋白(缺铁诱导蛋白C)由orf5(现称为idiC)编码,orf5是聚球藻PCC 7942铁响应idiB操纵子的一部分。20.5 kDa的IdiC蛋白具有一个推定的跨膜螺旋,属于硫氧还蛋白(TRX)样[2Fe-2S]铁氧化还原蛋白家族。IdiC与大肠杆菌NDH-1复合物的外周亚基NuoE具有最高的相似性。IdiC的表达在铁饥饿条件下以及生长后期增加,这代表了有利于光合循环和呼吸电子传递而非从水到NADP+的光合线性电子传递的生长条件。试图通过插入失活idiC基因产生了IdiC含量大幅降低的部分二倍体突变体(突变体MuD),但没有无IdiC的突变体。因此,在所用的实验条件下,IdiC似乎是聚球藻生存能力所必需的蛋白质。聚球藻野生型(WT)和突变体MuD的比较分析表明,在WT和MuD中,铁限制条件下反应中心蛋白PsbA和PsaA/B的量大幅减少。与WT相比,MuD以碳酸氢盐作为电子受体时具有更低的生长速率、叶绿素含量和光合放氧活性。免疫印迹分析还表明,在铁限制条件下生长时,与WT相比,MuD中IdiC和IdiB蛋白的量大幅减少。由于转录因子IdiB的减少,IdiA和IrpA的表达也降低。此外,尽管MuD和WT中的isiA mRNA水平相同,但MuD中的IsiA蛋白浓度低于WT。另一个显著差异是,与WT相比,在铁限制条件下,突变体MuD中铁氧化还原蛋白:NADP+氧化还原酶的表达较低。将讨论蛋白IdiC在光系统I周围的循环电子传递和/或呼吸电子传递中的可能功能。

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