CLSY docking to Pol IV requires a conserved domain critical for small RNA biogenesis and transposon silencing

Abstract Eukaryotes must balance the need for gene transcription by RNA polymerase II (Pol II) against the danger of mutations caused by transposable element (TE) proliferation. In plants, these gene expression and TE silencing activities are divided between different RNA polymerases. Specifically,...

Full description

Saved in:
Bibliographic Details
Main Authors: Luisa Felgines, Bart Rymen, Laura M. Martins, Guanghui Xu, Calvin Matteoli, Christophe Himber, Ming Zhou, Josh Eis, Ceyda Coruh, Marcel Böhrer, Lauriane Kuhn, Johana Chicher, Vijaya Pandey, Philippe Hammann, James Wohlschlegel, Florent Waltz, Julie A. Law, Todd Blevins
Format: Article
Language:English
Published: Nature Portfolio 2024-11-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-024-54268-0
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850107638687203328
author Luisa Felgines
Bart Rymen
Laura M. Martins
Guanghui Xu
Calvin Matteoli
Christophe Himber
Ming Zhou
Josh Eis
Ceyda Coruh
Marcel Böhrer
Lauriane Kuhn
Johana Chicher
Vijaya Pandey
Philippe Hammann
James Wohlschlegel
Florent Waltz
Julie A. Law
Todd Blevins
author_facet Luisa Felgines
Bart Rymen
Laura M. Martins
Guanghui Xu
Calvin Matteoli
Christophe Himber
Ming Zhou
Josh Eis
Ceyda Coruh
Marcel Böhrer
Lauriane Kuhn
Johana Chicher
Vijaya Pandey
Philippe Hammann
James Wohlschlegel
Florent Waltz
Julie A. Law
Todd Blevins
author_sort Luisa Felgines
collection DOAJ
description Abstract Eukaryotes must balance the need for gene transcription by RNA polymerase II (Pol II) against the danger of mutations caused by transposable element (TE) proliferation. In plants, these gene expression and TE silencing activities are divided between different RNA polymerases. Specifically, RNA polymerase IV (Pol IV), which evolved from Pol II, transcribes TEs to generate small interfering RNAs (siRNAs) that guide DNA methylation and block TE transcription by Pol II. While the Pol IV complex is recruited to TEs via SNF2-like CLASSY (CLSY) proteins, how Pol IV partners with the CLSYs remains unknown. Here, we identified a conserved CYC-YPMF motif that is specific to Pol IV and is positioned on the complex exterior. Furthermore, we found that this motif is essential for the co-purification of all four CLSYs with Pol IV, but that only one CLSY is present in any given Pol IV complex. These findings support a “one CLSY per Pol IV” model where the CYC-YPMF motif acts as a CLSY-docking site. Indeed, mutations in and around this motif phenocopy pol iv null and clsy quadruple mutants. Together, these findings provide structural and functional insights into a critical protein feature that distinguishes Pol IV from other RNA polymerases, allowing it to promote genome stability by targeting TEs for silencing.
format Article
id doaj-art-e8bc24794bde4fd99cb9297cd2c4e890
institution OA Journals
issn 2041-1723
language English
publishDate 2024-11-01
publisher Nature Portfolio
record_format Article
series Nature Communications
spelling doaj-art-e8bc24794bde4fd99cb9297cd2c4e8902025-08-20T02:38:32ZengNature PortfolioNature Communications2041-17232024-11-0115111610.1038/s41467-024-54268-0CLSY docking to Pol IV requires a conserved domain critical for small RNA biogenesis and transposon silencingLuisa Felgines0Bart Rymen1Laura M. Martins2Guanghui Xu3Calvin Matteoli4Christophe Himber5Ming Zhou6Josh Eis7Ceyda Coruh8Marcel Böhrer9Lauriane Kuhn10Johana Chicher11Vijaya Pandey12Philippe Hammann13James Wohlschlegel14Florent Waltz15Julie A. Law16Todd Blevins17Institut de Biologie Moléculaire des Plantes, CNRS, Université de StrasbourgInstitut de Biologie Moléculaire des Plantes, CNRS, Université de StrasbourgPlant Molecular and Cellular Biology Laboratory, Salk Institute for Biological StudiesPlant Molecular and Cellular Biology Laboratory, Salk Institute for Biological StudiesInstitut de Biologie Moléculaire des Plantes, CNRS, Université de StrasbourgInstitut de Biologie Moléculaire des Plantes, CNRS, Université de StrasbourgPlant Molecular and Cellular Biology Laboratory, Salk Institute for Biological StudiesPlant Molecular and Cellular Biology Laboratory, Salk Institute for Biological StudiesPlant Molecular and Cellular Biology Laboratory, Salk Institute for Biological StudiesInstitut de Biologie Moléculaire des Plantes, CNRS, Université de StrasbourgInstitut de Biologie Moléculaire et Cellulaire, CNRS, Plateforme Protéomique Strasbourg-EsplanadeInstitut de Biologie Moléculaire et Cellulaire, CNRS, Plateforme Protéomique Strasbourg-EsplanadeDepartment of Biological Chemistry, University of CaliforniaInstitut de Biologie Moléculaire et Cellulaire, CNRS, Plateforme Protéomique Strasbourg-EsplanadeDepartment of Biological Chemistry, University of CaliforniaBiozentrum, University of BaselPlant Molecular and Cellular Biology Laboratory, Salk Institute for Biological StudiesInstitut de Biologie Moléculaire des Plantes, CNRS, Université de StrasbourgAbstract Eukaryotes must balance the need for gene transcription by RNA polymerase II (Pol II) against the danger of mutations caused by transposable element (TE) proliferation. In plants, these gene expression and TE silencing activities are divided between different RNA polymerases. Specifically, RNA polymerase IV (Pol IV), which evolved from Pol II, transcribes TEs to generate small interfering RNAs (siRNAs) that guide DNA methylation and block TE transcription by Pol II. While the Pol IV complex is recruited to TEs via SNF2-like CLASSY (CLSY) proteins, how Pol IV partners with the CLSYs remains unknown. Here, we identified a conserved CYC-YPMF motif that is specific to Pol IV and is positioned on the complex exterior. Furthermore, we found that this motif is essential for the co-purification of all four CLSYs with Pol IV, but that only one CLSY is present in any given Pol IV complex. These findings support a “one CLSY per Pol IV” model where the CYC-YPMF motif acts as a CLSY-docking site. Indeed, mutations in and around this motif phenocopy pol iv null and clsy quadruple mutants. Together, these findings provide structural and functional insights into a critical protein feature that distinguishes Pol IV from other RNA polymerases, allowing it to promote genome stability by targeting TEs for silencing.https://doi.org/10.1038/s41467-024-54268-0
spellingShingle Luisa Felgines
Bart Rymen
Laura M. Martins
Guanghui Xu
Calvin Matteoli
Christophe Himber
Ming Zhou
Josh Eis
Ceyda Coruh
Marcel Böhrer
Lauriane Kuhn
Johana Chicher
Vijaya Pandey
Philippe Hammann
James Wohlschlegel
Florent Waltz
Julie A. Law
Todd Blevins
CLSY docking to Pol IV requires a conserved domain critical for small RNA biogenesis and transposon silencing
Nature Communications
title CLSY docking to Pol IV requires a conserved domain critical for small RNA biogenesis and transposon silencing
title_full CLSY docking to Pol IV requires a conserved domain critical for small RNA biogenesis and transposon silencing
title_fullStr CLSY docking to Pol IV requires a conserved domain critical for small RNA biogenesis and transposon silencing
title_full_unstemmed CLSY docking to Pol IV requires a conserved domain critical for small RNA biogenesis and transposon silencing
title_short CLSY docking to Pol IV requires a conserved domain critical for small RNA biogenesis and transposon silencing
title_sort clsy docking to pol iv requires a conserved domain critical for small rna biogenesis and transposon silencing
url https://doi.org/10.1038/s41467-024-54268-0
work_keys_str_mv AT luisafelgines clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT bartrymen clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT laurammartins clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT guanghuixu clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT calvinmatteoli clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT christophehimber clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT mingzhou clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT josheis clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT ceydacoruh clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT marcelbohrer clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT laurianekuhn clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT johanachicher clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT vijayapandey clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT philippehammann clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT jameswohlschlegel clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT florentwaltz clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT juliealaw clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing
AT toddblevins clsydockingtopolivrequiresaconserveddomaincriticalforsmallrnabiogenesisandtransposonsilencing