title: Interchangeability and distinct properties of bacterial Fe-S cluster assembly systems: functional replacement of the isc and suf operons in Escherichia coli with the nifSU-like operon from Helicobacter pylori.
abstract: The assembly of iron-sulfur (Fe-S) clusters, a key step in the post-translational maturation of Fe-S proteins, is mediated by a complex apparatus. In E. coli, this process involves two independent systems called ISC and SUF encoded by the iscSUA-hscBA-fdx gene cluster and sufABCDSE operon, respectively. Another system, termed NIF (nifSU), is required for the maturation of nitrogenase in nitrogen-fixing bacteria. We have developed a novel genetic system to gain further insight into these multi-component systems, and to determine how ISC, SUF and NIF might differ in their roles in Fe-S assembly. We have constructed an E. coli mutant lacking both the isc and suf operons, and this strain can only survive in the presence of a complementing plasmid. Using the plasmid replacement technique, we examined the isc and suf operons, and identified the genes essential for the function. Additionally, we have found that nifSU-like genes cloned from Helicobacter pylori are functionally exchangeable with the isc and suf operons. Thus, the NIF-like system participates in the maturation of a wide variety of Fe-S proteins. An increased ability of NIF to complement isc and suf loss was seen under anaerobic conditions. This may explain why the NIF system is only found in a limited number of bacterial species, and most other organisms prefer the ISC and/or SUF systems. While the differences between ISC and SUF were small with respect to the complementing activity, the SUF system appears to be more advantageous for bacterial growth in the presence of hydrogen peroxide.
authors.affiliation: Department of Biology, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043.
authors.forename: Umechiyo
authors.initials: U
authors.lastname: Tokumoto
authors.forename: Seiichi
authors.initials: S
authors.lastname: Kitamura
authors.forename: Keiichi
authors.initials: K
authors.lastname: Fukuyama
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Takahashi
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D016480:Helicobacter pylori / Q000235:genetics*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007501:Iron / Q000378:metabolism; D007506:Iron-Sulfur Proteins / Q000737:chemistry*; D008957:Models, Genetic; D005810:Multigene Family; D009154:Mutation; D009584:Nitrogen / Q000378:metabolism; D009591:Nitrogenase / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry; D009876:Operon; D018384:Oxidative Stress; D010641:Phenotype; D010957:Plasmids / Q000378:metabolism; D017382:Reactive Oxygen Species; D013997:Time Factors; D014157:Transcription Factors / Q000737:chemistry* / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D007506:Iron-Sulfur Proteins; C499695:IscR protein, E coli; D009841:Oligonucleotides; D017382:Reactive Oxygen Species; D014157:Transcription Factors; D006861:Hydrogen Peroxide; D007501:Iron; D009591:Nitrogenase; D009584:Nitrogen