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Stabilisation of the RirA [4Fe-4S] cluster results in loss of iron-sensing function

Chem Sci. 2023-08; 
Elizabeth Gray, Melissa Y Y Stewart, Libby Hanwell, Jason C Crack, Rebecca Devine, Clare E M Stevenson, Anne Volbeda, Andrew W B Johnston, Juan C Fontecilla-Camps, Matthew I Hutchings, Jonathan D Todd, Nick E Le Brun
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Gene Synthesis … rirA genes encoding the N8D and N8C variants of RirA were synthesised by Genscript and cloned using Ndel and BamH1 into the pET11a vector (Novagen) for protein overexpression … Get A Quote


RirA is a global iron regulator in diverse that belongs to the Rrf2 superfamily of transcriptional regulators, which can contain an iron-sulfur (Fe-S) cluster. Under iron-replete conditions, RirA contains a [4Fe-4S] cluster, enabling high-affinity binding to RirA-regulated operator sequences, thereby causing the repression of cellular iron uptake. Under iron deficiency, one of the cluster irons dissociates, generating an unstable [3Fe-4S] form that subsequently degrades to a [2Fe-2S] form and then to apo RirA, resulting in loss of high-affinity DNA-binding. The cluster is coordinated by three conserved cysteine residues and an unknown fourth ligand. Considering the lability of one of the irons and the resultin... More