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- W2028890624 abstract "Fe-S cluster, the nonheme-iron cofactor essential for the activity of many proteins, is incorporated into its target protein by an unknown mechanism. In Escherichia coli, genes in the OKFl-OBF2-iscS-iscU-iscA-hscB-hscA-fdx-ORF3 cluster (the isc gene cluster) should be involved in the assembly of the Fe-S cluster since its coexpression with the reporter ferredoxin (Fd) dramatically increases the production of holoFd [Nakamura, M., Saeki, K., and Takahashi, Y. (1999) J. Biochem. 126, 10–18]. In this study we addressed the functional roles of the proteins encoded by the isc gene cluster with respect to the assembly of Fe-S clusters in four reporter Fds. Plasmids were constructed in which eight ORFs in the we gene cluster were individually inactivated either by truncating the coding region or by introducing an oligonucleotide linker containing stop codons. By coexpressing these plasmids with reporter Fds, we show the iscS, iscA, hscA, and fdx genes to be required for the assembly of the Fe-S clusters. When these genes were absent from the coexpression plasmid, no overproduction was achieved in any reporter Fds examined. The inactivation of ORF2 and hscB had a partial but appreciable effect on the production of some Fds. Deletion of ORFl produced no difference from the coexpression with the intact isc gene cluster. We also examined coexpression using the fdx gene in the isc gene cluster as a reporter Fd and identified iscS, hscB, hscA, and ORF3 as being involved in the assembly of the [2Fe-2S] cluster in this protein. We propose a model in which the fdx gene product functions as an intermediate site for Fe-S cluster assembly." @default.
- W2028890624 created "2016-06-24" @default.
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- W2028890624 date "1999-11-01" @default.
- W2028890624 modified "2023-10-03" @default.
- W2028890624 title "Functional Assignment of the ORF2-iscS-iscU-iscA-hscB-hscA-fdx-0RF3 Gene Cluster Involved in the Assembly of Fe-S Clusters in Escherichia coli" @default.
- W2028890624 doi "https://doi.org/10.1093/oxfordjournals.jbchem.a022535" @default.
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