The Association Kinetics Encode the Light Dependence of Arabidopsis Phytochrome B Interactions

Titelangaben

Golonka, David ; Gerken, Uwe ; Köhler, Jürgen ; Möglich, Andreas:
The Association Kinetics Encode the Light Dependence of Arabidopsis Phytochrome B Interactions.
In: Journal of Molecular Biology. Bd. 432 (2020) Heft 16 . - S. 4327-4340.
ISSN 0022-2836
DOI: https://doi.org/10.1016/j.jmb.2020.06.001

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Abstract

Plant phytochromes enable vital adaptations to red and far-red light. At the molecular level, these responses are mediated by light-regulated interactions between phytochromes and partner proteins, foremost the phytochrome-interacting factors (PIF). Although known for decades, quantitative analyses of these interactions have long been sparse. To address this deficit, we here studied by an integrated fluorescence-spectroscopic approach the equilibrium and kinetics of Arabidopsis thaliana phytochrome B (AtPhyB) binding to a tetramerized PIF6 variant. Several readouts consistently showed the stringently light-regulated interaction to be little affected by PIF tetramerization. Analysis of the binding kinetics allowed the determination of bimolecular association and unimolecular dissociation rate constants as a function of light. Unexpectedly, the stronger affinity of AtPhyB under red light relative to far-red light is entirely due to accelerated association rather than decelerated dissociation. The association reaction under red light is highly efficient and only threefold slower than the diffusion limit. The present findings pertain equally to the analysis of signal transduction in plants and to the biotechnological application of phytochromes.