title: Phycoviolobilin formation and spectral tuning in the DXCF cyanobacteriochrome subfamily.
abstract: Phytochromes are red/far-red photosensory proteins that regulate adaptive responses to light via photoswitching of cysteine-linked linear tetrapyrrole (bilin) chromophores. The related cyanobacteriochromes (CBCRs) extend the photosensory range of the phytochrome superfamily to shorter wavelengths of visible light. CBCRs and phytochromes share a conserved Cys residue required for bilin attachment. In one CBCR subfamily, often associated with a blue/green photocycle, a second Cys lies within a conserved Asp-Xaa-Cys-Phe (DXCF) motif and is essential for the blue/green photocycle. Such DXCF CBCRs use isomerization of the phycocyanobilin (PCB) chromophore into the related phycoviolobilin (PVB) to shorten the conjugated system for sensing green light. We here use recombinant expression of individual CBCR domains in Escherichia coli to survey the DXCF subfamily from the cyanobacterium Nostoc punctiforme. We describe ten new photoreceptors with well-resolved photocycles and three additional photoproteins with overlapping dark-adapted and photoproduct states. We show that the ability of this subfamily to form PVB or retain PCB provides a powerful mechanism for tuning the photoproduct absorbance, with blue-absorbing dark states leading to a broad range of photoproducts absorbing teal, green, yellow, or orange light. Moreover, we use a novel green/teal CBCR that lacks the blue-absorbing dark state to demonstrate that PVB formation requires the DXCF Cys residue. Our results demonstrate that this subfamily exhibits much more spectral diversity than had been previously appreciated.
authors.affiliation: Department of Molecular and Cellular Biology, University of California, Davis, California 95616, United States.
authors.forename: Nathan C
authors.initials: NC
authors.lastname: Rockwell
authors.forename: Shelley S
authors.initials: SS
authors.lastname: Martin
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Gulevich
authors.forename: J Clark
authors.initials: JC
authors.lastname: Lagarias
mesh_terms: D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods; D017124:Conserved Sequence; D000458:Cyanobacteria / Q000378:metabolism*; D003545:Cysteine / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008027:Light; D008956:Models, Chemical; D009154:Mutation; D046937:Nostoc / Q000235:genetics / Q000378:metabolism*; D010777:Photochemistry / Q000379:methods; D020283:Photoreceptors, Microbial / Q000737:chemistry*; D052980:Phycobilins / Q000737:chemistry*; D013696:Temperature; D013997:Time Factors
chemical_list: D020283:Photoreceptors, Microbial; D052980:Phycobilins; C526232:phycoviolobilin; D003545:Cysteine