TY - JOUR
T1 - Deconstructing and repurposing the light-regulated interplay between Arabidopsis phytochromes and interacting factors
AU - Golonka, David
AU - Fischbach, Patrick
AU - Jena, Siddhartha G.
AU - Kleeberg, Julius R.W.
AU - Essen, Lars Oliver
AU - Toettcher, Jared E.
AU - Zurbriggen, Matias D.
AU - Möglich, Andreas
N1 - Funding Information:
We thank members of our laboratories for support and comments; Dr. J. Casal for fruitful discussion; Dr. B. Höcker for chromatography use; Dr. T. Scheibel for ultracentrifuge use; and Dr. M. Hörner and Dr. W. Weber for generously providing PhyB expression plasmids. Funding through the Deutsche Forschungsgemeinschaft (grants MO2192/7-1 to A. M., ZU259/2-1 to M.D.Z., and ES152/16 to L.-O.E.), under Germany’s Excellence Strategy CEPLAS EXC2048/1 (to M.D.Z., ID 390686111), by NIH grant DP2EB024247 (to J.E.T.), by NIH training grant T32GM007388 (to S.G.J.), and by the European Commission – Research Executive Agency (H2020 Future and Emerging Technologies (FET-Open) Project ID 801041 CyGenTiG to M.D.Z.) is appreciated. J.R.W.K. gratefully acknowledges support through the ENB program “Biological Physics.”
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - Phytochrome photoreceptors mediate adaptive responses of plants to red and far-red light. These responses generally entail light-regulated association between phytochromes and other proteins, among them the phytochrome-interacting factors (PIF). The interaction with Arabidopsis thaliana phytochrome B (AtPhyB) localizes to the bipartite APB motif of the A. thaliana PIFs (AtPIF). To address a dearth of quantitative interaction data, we construct and analyze numerous AtPIF3/6 variants. Red-light-activated binding is predominantly mediated by the APB N-terminus, whereas the C-terminus modulates binding and underlies the differential affinity of AtPIF3 and AtPIF6. We identify AtPIF variants of reduced size, monomeric or homodimeric state, and with AtPhyB affinities between 10 and 700 nM. Optogenetically deployed in mammalian cells, the AtPIF variants drive light-regulated gene expression and membrane recruitment, in certain cases reducing basal activity and enhancing regulatory response. Moreover, our results provide hitherto unavailable quantitative insight into the AtPhyB:AtPIF interaction underpinning vital light-dependent responses in plants.
AB - Phytochrome photoreceptors mediate adaptive responses of plants to red and far-red light. These responses generally entail light-regulated association between phytochromes and other proteins, among them the phytochrome-interacting factors (PIF). The interaction with Arabidopsis thaliana phytochrome B (AtPhyB) localizes to the bipartite APB motif of the A. thaliana PIFs (AtPIF). To address a dearth of quantitative interaction data, we construct and analyze numerous AtPIF3/6 variants. Red-light-activated binding is predominantly mediated by the APB N-terminus, whereas the C-terminus modulates binding and underlies the differential affinity of AtPIF3 and AtPIF6. We identify AtPIF variants of reduced size, monomeric or homodimeric state, and with AtPhyB affinities between 10 and 700 nM. Optogenetically deployed in mammalian cells, the AtPIF variants drive light-regulated gene expression and membrane recruitment, in certain cases reducing basal activity and enhancing regulatory response. Moreover, our results provide hitherto unavailable quantitative insight into the AtPhyB:AtPIF interaction underpinning vital light-dependent responses in plants.
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U2 - 10.1038/s42003-019-0687-9
DO - 10.1038/s42003-019-0687-9
M3 - Article
C2 - 31925272
AN - SCOPUS:85075954158
SN - 2399-3642
VL - 2
JO - Communications Biology
JF - Communications Biology
IS - 1
M1 - 448
ER -