RRC ID 35641
著者 Hidese R, Mihara H, Kurihara T, Esaki N.
タイトル Global identification of genes affecting iron-sulfur cluster biogenesis and iron homeostasis.
ジャーナル J Bacteriol
Abstract Iron-sulfur (Fe-S) clusters are ubiquitous cofactors that are crucial for many physiological processes in all organisms. In Escherichia coli, assembly of Fe-S clusters depends on the activity of the iron-sulfur cluster (ISC) assembly and sulfur mobilization (SUF) apparatus. However, the underlying molecular mechanisms and the mechanisms that control Fe-S cluster biogenesis and iron homeostasis are still poorly defined. In this study, we performed a global screen to identify the factors affecting Fe-S cluster biogenesis and iron homeostasis using the Keio collection, which is a library of 3,815 single-gene E. coli knockout mutants. The approach was based on radiolabeling of the cells with [2-(14)C]dihydrouracil, which entirely depends on the activity of an Fe-S enzyme, dihydropyrimidine dehydrogenase. We identified 49 genes affecting Fe-S cluster biogenesis and/or iron homeostasis, including 23 genes important only under microaerobic/anaerobic conditions. This study defines key proteins associated with Fe-S cluster biogenesis and iron homeostasis, which will aid further understanding of the cellular mechanisms that coordinate the processes. In addition, we applied the [2-(14)C]dihydrouracil-labeling method to analyze the role of amino acid residues of an Fe-S cluster assembly scaffold (IscU) as a model of the Fe-S cluster assembly apparatus. The analysis showed that Cys37, Cys63, His105, and Cys106 are essential for the function of IscU in vivo, demonstrating the potential of the method to investigate in vivo function of proteins involved in Fe-S cluster assembly.
巻・号 196(6)
ページ 1238-49
公開日 2014-3-1
DOI 10.1128/JB.01160-13
PII JB.01160-13
PMID 24415728
PMC PMC3957717
MeSH Escherichia coli / genetics* Escherichia coli / metabolism* Escherichia coli Proteins / genetics* Escherichia coli Proteins / metabolism* Gene Knockout Techniques Homeostasis Iron / metabolism* Isotope Labeling Metabolic Networks and Pathways / genetics* Sulfur / metabolism* Uracil / analogs & derivatives Uracil / metabolism
IF 3.006
引用数 8
WOS 分野 MICROBIOLOGY
リソース情報
原核生物(大腸菌) ME9062(BW25113) Keio collection