Structural basis for the photoconversion of a phytochrome to the activated Pfr form

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Abstract

Phytochromes are a collection of bilin-containing photoreceptors that regulate numerous photoresponses in plants and microorganisms through their ability to photointerconvert between a red-light-absorbing, ground state (Pr) and a far-red-light-absorbing, photoactivated state (Pfr)1,2. Although the structures of several phytochromes as Pr have been determined3,4,5,6,7, little is known about the structure of Pfr and how it initiates signalling. Here we describe the three-dimensional solution structure of the bilin-binding domain as Pfr, using the cyanobacterial phytochrome from Synechococcus OSB′. Contrary to predictions, light-induced rotation of the A pyrrole ring but not the D ring is the primary motion of the chromophore during photoconversion. Subsequent rearrangements within the protein then affect intradomain and interdomain contact sites within the phytochrome dimer. On the basis of our models, we propose that phytochromes act by propagating reversible light-driven conformational changes in the bilin to altered contacts between the adjacent output domains, which in most phytochromes direct differential phosphotransfer.

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Figure 1: Three-dimensional overlay of SyB -Cph1(GAF) Pr and Pfr solution structures.
Figure 2: Rotation of the A ring of the PCB chromophore during Pr to Pfr photoconversion.
Figure 3: Light-driven conformational changes for amino acids surrounding the chromophore.
Figure 4: Conformational rearrangement of Asp 86, Tyr 142 and Phe 82 during photoconversion from Pr to Pfr.

Accession codes

Primary accessions

Protein Data Bank

Data deposits

Atomic coordinates and structures for Pr and Pfr are deposited in the Protein Data Bank under accession codes 2KOI and 2KLI, respectively.

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Acknowledgements

We thank K. T. Forest and W. M. Westler for technical advice. This work was supported by a grant from the US National Science Foundation (R.D.V.) and a postdoctoral fellowship from the American Heart Association (A.T.U.). C.C.C. was supported by the U.S. National Institutes of Health. This study was a collaboration with the National Magnetic Resonance Facility at Madison, which is supported by the US National Institute of Health.

Author Contributions R.D.V. and A.T.U. initiated the collaboration. A.T.U. and J.Z. purified the chromoproteins and A.T.U. characterized the samples. M.R. provided the isotopically labelled ALA. G.C. collected the NMR spectra and solved the NMR structures with C.C.C. and J.L.M. All authors interpreted the three-dimensional structures. R.D.V., A.T.U., C.C.C. and G.C. prepared the manuscript, tables and figures.

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Correspondence to Richard D. Vierstra.

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This file contains Supplementary Table 1, Supplementary Figures 1-7 with Legends and Supplementary References. The y-axis of Supplementary Fig. 4e was corrected on January 25 2010. (PDF 2162 kb)

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Ulijasz, A., Cornilescu, G., Cornilescu, C. et al. Structural basis for the photoconversion of a phytochrome to the activated Pfr form. Nature 463, 250–254 (2010) doi:10.1038/nature08671

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