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Revisiting chromatin binding of the Arabidopsis UV-B photoreceptor UVR8

BACKGROUND: Plants perceive UV-B through the UV RESISTANCE LOCUS 8 (UVR8) photoreceptor and UVR8 activation leads to changes in gene expression such as those associated with UV-B acclimation and stress tolerance. Albeit functionally unrelated, UVR8 shows some homology with RCC1 (Regulator of Chromat...

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Autores principales: Binkert, Melanie, Crocco, Carlos D., Ekundayo, Babatunde, Lau, Kelvin, Raffelberg, Sarah, Tilbrook, Kimberley, Yin, Ruohe, Chappuis, Richard, Schalch, Thomas, Ulm, Roman
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750278/
https://www.ncbi.nlm.nih.gov/pubmed/26864020
http://dx.doi.org/10.1186/s12870-016-0732-5
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author Binkert, Melanie
Crocco, Carlos D.
Ekundayo, Babatunde
Lau, Kelvin
Raffelberg, Sarah
Tilbrook, Kimberley
Yin, Ruohe
Chappuis, Richard
Schalch, Thomas
Ulm, Roman
author_facet Binkert, Melanie
Crocco, Carlos D.
Ekundayo, Babatunde
Lau, Kelvin
Raffelberg, Sarah
Tilbrook, Kimberley
Yin, Ruohe
Chappuis, Richard
Schalch, Thomas
Ulm, Roman
author_sort Binkert, Melanie
collection PubMed
description BACKGROUND: Plants perceive UV-B through the UV RESISTANCE LOCUS 8 (UVR8) photoreceptor and UVR8 activation leads to changes in gene expression such as those associated with UV-B acclimation and stress tolerance. Albeit functionally unrelated, UVR8 shows some homology with RCC1 (Regulator of Chromatin Condensation 1) proteins from non-plant organisms at the sequence level. These proteins act as guanine nucleotide exchange factors for Ran GTPases and bind chromatin via histones. Subsequent to the revelation of this sequence homology, evidence was presented showing that UVR8 activity involves interaction with chromatin at the loci of some target genes through histone binding. This suggested a UVR8 mode-of-action intimately and directly linked with gene transcription. However, several aspects of UVR8 chromatin association remained undefined, namely the impact of UV-B on the process and how UVR8 chromatin association related to the transcription factor ELONGATED HYPOCOTYL 5 (HY5), which is important for UV-B signalling and has overlapping chromatin targets. Therefore, we have investigated UVR8 chromatin association in further detail. RESULTS: Unlike the claims of previous studies, our chromatin immunoprecipitation (ChIP) experiments do not confirm UVR8 chromatin association. In contrast to human RCC1, recombinant UVR8 also does not bind nucleosomes in vitro. Moreover, fusion of a VP16 activation domain to UVR8 did not alter expression of proposed UVR8 target genes in transient gene expression assays. Finally, comparison of the Drosophila DmRCC1 and the Arabidopsis UVR8 crystal structures revealed that critical histone- and DNA-interaction residues apparent in DmRCC1 are not conserved in UVR8. CONCLUSION: This has led us to conclude that the cellular activity of UVR8 likely does not involve its specific binding to chromatin at target genes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12870-016-0732-5) contains supplementary material, which is available to authorized users.
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spelling pubmed-47502782016-02-12 Revisiting chromatin binding of the Arabidopsis UV-B photoreceptor UVR8 Binkert, Melanie Crocco, Carlos D. Ekundayo, Babatunde Lau, Kelvin Raffelberg, Sarah Tilbrook, Kimberley Yin, Ruohe Chappuis, Richard Schalch, Thomas Ulm, Roman BMC Plant Biol Research Article BACKGROUND: Plants perceive UV-B through the UV RESISTANCE LOCUS 8 (UVR8) photoreceptor and UVR8 activation leads to changes in gene expression such as those associated with UV-B acclimation and stress tolerance. Albeit functionally unrelated, UVR8 shows some homology with RCC1 (Regulator of Chromatin Condensation 1) proteins from non-plant organisms at the sequence level. These proteins act as guanine nucleotide exchange factors for Ran GTPases and bind chromatin via histones. Subsequent to the revelation of this sequence homology, evidence was presented showing that UVR8 activity involves interaction with chromatin at the loci of some target genes through histone binding. This suggested a UVR8 mode-of-action intimately and directly linked with gene transcription. However, several aspects of UVR8 chromatin association remained undefined, namely the impact of UV-B on the process and how UVR8 chromatin association related to the transcription factor ELONGATED HYPOCOTYL 5 (HY5), which is important for UV-B signalling and has overlapping chromatin targets. Therefore, we have investigated UVR8 chromatin association in further detail. RESULTS: Unlike the claims of previous studies, our chromatin immunoprecipitation (ChIP) experiments do not confirm UVR8 chromatin association. In contrast to human RCC1, recombinant UVR8 also does not bind nucleosomes in vitro. Moreover, fusion of a VP16 activation domain to UVR8 did not alter expression of proposed UVR8 target genes in transient gene expression assays. Finally, comparison of the Drosophila DmRCC1 and the Arabidopsis UVR8 crystal structures revealed that critical histone- and DNA-interaction residues apparent in DmRCC1 are not conserved in UVR8. CONCLUSION: This has led us to conclude that the cellular activity of UVR8 likely does not involve its specific binding to chromatin at target genes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12870-016-0732-5) contains supplementary material, which is available to authorized users. BioMed Central 2016-02-11 /pmc/articles/PMC4750278/ /pubmed/26864020 http://dx.doi.org/10.1186/s12870-016-0732-5 Text en © Binkert et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Binkert, Melanie
Crocco, Carlos D.
Ekundayo, Babatunde
Lau, Kelvin
Raffelberg, Sarah
Tilbrook, Kimberley
Yin, Ruohe
Chappuis, Richard
Schalch, Thomas
Ulm, Roman
Revisiting chromatin binding of the Arabidopsis UV-B photoreceptor UVR8
title Revisiting chromatin binding of the Arabidopsis UV-B photoreceptor UVR8
title_full Revisiting chromatin binding of the Arabidopsis UV-B photoreceptor UVR8
title_fullStr Revisiting chromatin binding of the Arabidopsis UV-B photoreceptor UVR8
title_full_unstemmed Revisiting chromatin binding of the Arabidopsis UV-B photoreceptor UVR8
title_short Revisiting chromatin binding of the Arabidopsis UV-B photoreceptor UVR8
title_sort revisiting chromatin binding of the arabidopsis uv-b photoreceptor uvr8
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4750278/
https://www.ncbi.nlm.nih.gov/pubmed/26864020
http://dx.doi.org/10.1186/s12870-016-0732-5
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