Cargando…

PRC2 direct transfer from G-quadruplex RNA to dsDNA has implications for RNA-binding chromatin modifiers

The chromatin-modifying enzyme, Polycomb Repressive Complex 2 (PRC2), deposits the H3K27me3 epigenetic mark to negatively regulate expression at numerous target genes, and this activity has been implicated in embryonic development, cell differentiation, and various cancers. A biological role for RNA...

Descripción completa

Detalles Bibliográficos
Autores principales: Hemphill, Wayne O., Fenske, Regan, Gooding, Anne R., Cech, Thomas R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10266057/
https://www.ncbi.nlm.nih.gov/pubmed/37252986
http://dx.doi.org/10.1073/pnas.2220528120
_version_ 1785058664673968128
author Hemphill, Wayne O.
Fenske, Regan
Gooding, Anne R.
Cech, Thomas R.
author_facet Hemphill, Wayne O.
Fenske, Regan
Gooding, Anne R.
Cech, Thomas R.
author_sort Hemphill, Wayne O.
collection PubMed
description The chromatin-modifying enzyme, Polycomb Repressive Complex 2 (PRC2), deposits the H3K27me3 epigenetic mark to negatively regulate expression at numerous target genes, and this activity has been implicated in embryonic development, cell differentiation, and various cancers. A biological role for RNA binding in regulating PRC2 histone methyltransferase activity is generally accepted, but the nature and mechanism of this relationship remains an area of active investigation. Notably, many in vitro studies demonstrate that RNA inhibits PRC2 activity on nucleosomes through mutually antagonistic binding, while some in vivo studies indicate that PRC2’s RNA-binding activity is critical for facilitating its biological function(s). Here we use biochemical, biophysical, and computational approaches to interrogate PRC2’s RNA and DNA-binding kinetics. Our findings demonstrate that PRC2-polynucleotide dissociation rates are dependent on the concentration of free ligand, indicating the potential for direct transfer between nucleic acid ligands without a free-enzyme intermediate. Direct transfer explains the variation in previously reported dissociation kinetics, allows reconciliation of prior in vitro and in vivo studies, and expands the potential mechanisms of RNA-mediated PRC2 regulation. Moreover, simulations indicate that such a direct transfer mechanism could be obligatory for RNA to recruit proteins to chromatin.
format Online
Article
Text
id pubmed-10266057
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher National Academy of Sciences
record_format MEDLINE/PubMed
spelling pubmed-102660572023-06-15 PRC2 direct transfer from G-quadruplex RNA to dsDNA has implications for RNA-binding chromatin modifiers Hemphill, Wayne O. Fenske, Regan Gooding, Anne R. Cech, Thomas R. Proc Natl Acad Sci U S A Biological Sciences The chromatin-modifying enzyme, Polycomb Repressive Complex 2 (PRC2), deposits the H3K27me3 epigenetic mark to negatively regulate expression at numerous target genes, and this activity has been implicated in embryonic development, cell differentiation, and various cancers. A biological role for RNA binding in regulating PRC2 histone methyltransferase activity is generally accepted, but the nature and mechanism of this relationship remains an area of active investigation. Notably, many in vitro studies demonstrate that RNA inhibits PRC2 activity on nucleosomes through mutually antagonistic binding, while some in vivo studies indicate that PRC2’s RNA-binding activity is critical for facilitating its biological function(s). Here we use biochemical, biophysical, and computational approaches to interrogate PRC2’s RNA and DNA-binding kinetics. Our findings demonstrate that PRC2-polynucleotide dissociation rates are dependent on the concentration of free ligand, indicating the potential for direct transfer between nucleic acid ligands without a free-enzyme intermediate. Direct transfer explains the variation in previously reported dissociation kinetics, allows reconciliation of prior in vitro and in vivo studies, and expands the potential mechanisms of RNA-mediated PRC2 regulation. Moreover, simulations indicate that such a direct transfer mechanism could be obligatory for RNA to recruit proteins to chromatin. National Academy of Sciences 2023-05-30 2023-06-06 /pmc/articles/PMC10266057/ /pubmed/37252986 http://dx.doi.org/10.1073/pnas.2220528120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by/4.0/This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY) (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Biological Sciences
Hemphill, Wayne O.
Fenske, Regan
Gooding, Anne R.
Cech, Thomas R.
PRC2 direct transfer from G-quadruplex RNA to dsDNA has implications for RNA-binding chromatin modifiers
title PRC2 direct transfer from G-quadruplex RNA to dsDNA has implications for RNA-binding chromatin modifiers
title_full PRC2 direct transfer from G-quadruplex RNA to dsDNA has implications for RNA-binding chromatin modifiers
title_fullStr PRC2 direct transfer from G-quadruplex RNA to dsDNA has implications for RNA-binding chromatin modifiers
title_full_unstemmed PRC2 direct transfer from G-quadruplex RNA to dsDNA has implications for RNA-binding chromatin modifiers
title_short PRC2 direct transfer from G-quadruplex RNA to dsDNA has implications for RNA-binding chromatin modifiers
title_sort prc2 direct transfer from g-quadruplex rna to dsdna has implications for rna-binding chromatin modifiers
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10266057/
https://www.ncbi.nlm.nih.gov/pubmed/37252986
http://dx.doi.org/10.1073/pnas.2220528120
work_keys_str_mv AT hemphillwayneo prc2directtransferfromgquadruplexrnatodsdnahasimplicationsforrnabindingchromatinmodifiers
AT fenskeregan prc2directtransferfromgquadruplexrnatodsdnahasimplicationsforrnabindingchromatinmodifiers
AT goodinganner prc2directtransferfromgquadruplexrnatodsdnahasimplicationsforrnabindingchromatinmodifiers
AT cechthomasr prc2directtransferfromgquadruplexrnatodsdnahasimplicationsforrnabindingchromatinmodifiers