期刊
BIOCHEMICAL JOURNAL
卷 420, 期 -, 页码 463-472出版社
PORTLAND PRESS LTD
DOI: 10.1042/BJ20090206
关键词
aconitase; branched-chain amino acid (BCAA); dihydroxyacid dehydrogenase; iron-sulfur clusters; IscA/SufA paralogue; thiamin
资金
- Science and Technology Key Program of Zhejiang Province, China [2006C14025]
- Natural Science Foundation of Zhejiang Province, China [Y2081075, Y507233]
- United States National Sciences Foundation [MCB-0416537]
- National Institutes of Health [CA107494]
IscA/SufA paralogues are the members of the iron-sulfur cluster-assembly machinery in Escherichia coli. Whereas deletion of either IscA or SufA has only a mild effect on cell growth, deletion of both IscA and SufA results in a null-growth phenotype in minimal medium under aerobic growth conditions. Here we report that cell growth of the iscA/sufA double mutant (E. coli strain in which both iscA and sufA had been in-frame-deleted) call be partially restored by supplementing with BCAAs (branched-chain amino acids) and thiamin. We further demonstrate that deletion of IscA/SufA paralogues blocks the [4Fe-4S] cluster assembly in IlvD (dihydroxyacid dehydratase) of the BCAA biosynthetic pathway in E. coli cells under aerobic conditions and that addition of the iron-bound IscA/SufA efficiently promotes the [4Fe-4S] cluster assembly in IlvD and restores the enzyme activity in vitro, Suggesting that IscA/SufA may act as an iron donor for the [4Fe-4S] cluster assembly Under aerobic conditions. Additional studies reveal that IscA/SudA are also required for the [4Fe-4S] cluster assembly in enzyme ThiC of the thiamin-biosynthetic pathway, aconitase B of the citrate acid cycle and endonuclease III of the DNA-base-excision-repair pathway in E. coli under aerobic conditions. Nevertheless, deletion of IscA/SufA does not significantly affect the [2Fe-2S] cluster assembly in the redox transcription factor SoxR, ferredoxin and the siderophore-iron reductase FhuF. The results suggest that the biogenesis of the [4Fe-4S] clusters and the [2Fe-2S] Clusters may have distinct pathways and that IscA/SufA paralogues are essential for the [4Fe-4S] cluster assembly, but are dispensable for the [2Fe-2S] Cluster assembly in E. coli under aerobic conditions.
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