FurC Regulates Expression of zupT for the Central Zinc Importer ZupT of Cupriavidus metallidurans

FurC Regulates Expression of zupT for the Central Zinc Importer ZupT of Cupriavidus metallidurans
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FurC 调节 Curiavidus Metallidurans 中央锌输入蛋白 ZupT 的表达

DOI:
10.1128/jb.01713-14
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发表时间:
2014
影响因子:
3.2
通讯作者:
Nies DH
Nies DH
中科院分区:
生物学3区
文献类型:
--
作者:
Schmidt C;Schwarzenberger C;Grosse C;Nies DH

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锌进口商ZupT是需要锌的有效分配锌依赖蛋白质的金属抗性细菌嗜铜菌metallidurans,但不是锌importper本身。zupT的表达在锌饥饿条件下上调。C. metallidurans包含有资格作为thezupTregulator候选者的Fur家族的三个成员。在Δ furC突变体中,azupT-lacZ报告基因融合体的表达强烈上调,但在Δ furA或Δ furB突变体中不上调。在所有三种Δfurmutants中,过渡金属输入基因(pitA、corA 1、corA 2和corA 3)的表达在这种模式下没有改变,但在锌浓度升高的条件下,它们仍然下调,表明存在另一种锌依赖性调节因子。FurA是C铁代谢的核心调节因子。metallidurans,和furA组成型表达的条件下测试。在铁饥饿条件下,furB的表达上调,ΔfurBΔ furC双突变体的表型表明,FurB可能是一种连接一般金属和铁稳态的铁饥饿Fur。FurC被纯化为Strep标记的蛋白质,并阻碍了zupT上游DNA片段的电泳迁移率。FurC与该操作区域的结合受到锌离子和EDTA的存在的影响。因此,FurC是C.当有足够的锌存在时,它抑制了中心锌输入者ZupT的合成。
The zinc importer ZupT is required for the efficient allocation of zinc to zinc-dependent proteins in the metal-resistant bacterium Cupriavidus metallidurans but not for zinc importper se. The expression ofzupTis upregulated under conditions of zinc starvation. C. metallidurans contains three members of the Fur family of regulators that qualify as candidates for thezupTregulator. The expression of azupT-lacZreporter gene fusion was strongly upregulated in a ΔfurCmutant but not in a ΔfurAor ΔfurBmutant. Expression of the genes for transition-metal importers (pitA,corA1,corA2, andcorA3) was not changed in this pattern in all three Δfurmutants, but they were still downregulated under conditions of elevated zinc concentrations, indicating the presence of another zinc-dependent regulator. FurA was a central regulator of the iron metabolism in C. metallidurans, andfurAwas constitutively expressed under the conditions tested. Expression offurBwas upregulated under conditions of iron starvation, and FurB could be an iron starvation Fur connecting general metal and iron homeostasis, as indicated by the phenotype of a ΔfurBΔfurCdouble mutant. FurC was purified as a Strep-tagged protein and retarded the electrophoretic mobility of a DNA fragment upstream ofzupT. Binding of FurC to this operator region was influenced by the presence of zinc ions and EDTA. Thus, FurC is the main zinc uptake regulator (Zur) of C. metallidurans and represses synthesis of the central zinc importer ZupT when sufficient zinc is present.
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