title: The E. coli monothiol glutaredoxin GrxD forms homodimeric and heterodimeric FeS cluster containing complexes.
abstract: Monothiol glutaredoxins (mono-Grx) represent a highly evolutionarily conserved class of proteins present in organisms ranging from prokaryotes to humans. Mono-Grxs have been implicated in iron sulfur (FeS) cluster biosynthesis as potential scaffold proteins and in iron homeostasis via an FeS-containing complex with Fra2p (homologue of E. coli BolA) in yeast and are linked to signal transduction in mammalian systems. However, the function of the mono-Grx in prokaryotes and the nature of an interaction with BolA-like proteins have not been established. Recent genome-wide screens for E. coli genetic interactions reported the synthetic lethality (combination of mutations leading to cell death; mutation of only one of these genes does not) of a grxD mutation when combined with strains defective in FeS cluster biosynthesis (isc operon) functions [Butland, G., et al. (2008) Nature Methods 5, 789-795]. These data connected the only E. coli mono-Grx, GrxD to a potential role in FeS cluster biosynthesis. We investigated GrxD to uncover the molecular basis of this synthetic lethality and observed that GrxD can form FeS-bound homodimeric and BolA containing heterodimeric complexes. These complexes display substantially different spectroscopic and functional properties, including the ability to act as scaffold proteins for intact FeS cluster transfer to the model [2Fe-2S] acceptor protein E. coli apo-ferredoxin (Fdx), with the homodimer being significantly more efficient. In this work, we functionally dissect the potential cellular roles of GrxD as a component of both homodimeric and heterodimeric complexes to ultimately uncover if either of these complexes performs functions linked to FeS cluster biosynthesis.
authors: Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
authors: N
authors: N
authors: Yeung
authors: B
authors: B
authors: Gold
authors: N L
authors: NL
authors: Liu
authors: R
authors: R
authors: Prathapam
authors: H J
authors: HJ
authors: Sterling
authors: E R
authors: ER
authors: Willams
authors: G
authors: G
authors: Butland
mesh_terms: D002850:Chromatography, Gel / Q000379:methods; D002942:Circular Dichroism / Q000379:methods; D019281:Dimerization; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D054477:Glutaredoxins / Q000737:chemistry*; D006801:Humans; D007501:Iron / Q000737:chemistry; D007506:Iron-Sulfur Proteins / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D008957:Models, Genetic; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D013053:Spectrophotometry / Q000379:methods; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
chemical_list: D029968:Escherichia coli Proteins; D054477:Glutaredoxins; C568707:GrxD protein, E coli; D007506:Iron-Sulfur Proteins; D007501:Iron