PMID:21295582

Citation	Greenwell, R, Nam, TW and Donohue, TJ (2011) Features of Rhodobacter sphaeroides ChrR required for stimuli to promote the dissociation of σ(E)/ChrR complexes. J. Mol. Biol. 407:477-91
Abstract	In the photosynthetic bacterium Rhodobacter sphaeroides, a transcriptional response to the reactive oxygen species singlet oxygen ((1)O(2)) is mediated by ChrR, a zinc metalloprotein that binds to and inhibits the activity of the alternative σ factor σ(E). We provide evidence that (1)O(2) promotes the dissociation of σ(E) from ChrR to activate transcription in vivo. To identify what is required for (1)O(2) to promote the dissociation of σ(E)/ChrR complexes, we analyzed the in vivo properties of variant ChrR proteins with amino acid changes in conserved residues of the C-terminal cupin-like domain (ChrR-CLD). We found that (1)O(2) was unable to promote the detectable dissociation of σ(E)/ChrR complexes when the ChrR-CLD zinc ligands (His141, His143, Glu147, and His177) were substituted with alanine, even though individual substitutions caused a 2-fold to 10-fold decrease in zinc affinity for this domain relative to that for wild-type ChrR (K(d)∼4.6×10(-)(10) M). We conclude that the side chains of these invariant residues play a crucial role in the response to (1)O(2). Additionally, we found that cells containing variant ChrR proteins with single amino acid substitutions at Cys187 or Cys189 exhibited σ(E) activity similar to those containing wild-type ChrR when exposed to (1)O(2), suggesting that these thiol side chains are not required for (1)O(2) to induce σ(E) activity in vivo. Finally, we found that the same aspects of R. sphaeroides ChrR needed for a response to (1)O(2) are required for the dissociation of σ(E)/ChrR complexes in the presence of the organic hydroperoxide t-butyl hydroperoxide.
Links	PubMed PMC3061837 Online version:10.1016/j.jmb.2011.01.055
Keywords	Amino Acid Substitution; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Coenzymes/metabolism; Gene Expression Regulation; Models, Molecular; Mutagenesis, Site-Directed; Mutant Proteins/genetics; Mutant Proteins/metabolism; Protein Binding; Protein Structure, Quaternary; Rhodobacter sphaeroides/genetics; Rhodobacter sphaeroides/metabolism; Sigma Factor/metabolism; Singlet Oxygen/metabolism; Transcription Factors/genetics; Transcription Factors/metabolism; Transcription, Genetic; Zinc/metabolism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
RHOS4:CHRR		GO:0016987: sigma factor activity	ECO:0000314:		F		Figure 2,3, and 4	complete CACAO 8980

References

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