Altered interactions within FY/AtCPSF complexes required for Arabidopsis FCA-mediated chromatin silencing

TY - JOUR

T1 - Altered interactions within FY/AtCPSF complexes required for Arabidopsis FCA-mediated chromatin silencing

AU - Manzano, David

AU - Marquardt, Sebastian

AU - Jones, Alexandra M E

AU - Bäurle, Isabel

AU - Liu, Fuquan

AU - Dean, Caroline

PY - 2009

Y1 - 2009

N2 - The role of RNA metabolism in chromatin silencing is now widely recognized. We have studied the Arabidopsis RNA-binding protein FCA that down-regulates an endogenous floral repressor gene through a chromatin mechanism involving histone demethylase activity. This mechanism needs FCA to interact with an RNA 3' processing/polyadenylation factor (FY/Pfs2p), but the subsequent events leading to chromatin changes are unknown. Here, we show that this FCA-FY interaction is required for general chromatin silencing roles where hairpin transgenes induce DNA methylation of an endogenous gene. We also show 2 conserved RNA processing factors, AtCPSF100 and AtCPSF160, but not FCA, are stably associated with FY in vivo and form a range of different-sized complexes. A hypomorphic fy allele producing a shorter protein, able to provide some FY functions but unable to interact with FCA, reduces abundance of some of the larger MW complexes. Suppressor mutants, which specifically disrupt the FY motif through which FCA interacts, also lacked these larger complexes. Our data support a model whereby FCA, perhaps after recognition of a specific RNA feature, transiently interacts with FY, an integral component of the canonical RNA 3' processing machinery, changing the interactions of the different RNA processing components. These altered interactions would appear to be a necessary step in this RNA-mediated chromatin silencing.

AB - The role of RNA metabolism in chromatin silencing is now widely recognized. We have studied the Arabidopsis RNA-binding protein FCA that down-regulates an endogenous floral repressor gene through a chromatin mechanism involving histone demethylase activity. This mechanism needs FCA to interact with an RNA 3' processing/polyadenylation factor (FY/Pfs2p), but the subsequent events leading to chromatin changes are unknown. Here, we show that this FCA-FY interaction is required for general chromatin silencing roles where hairpin transgenes induce DNA methylation of an endogenous gene. We also show 2 conserved RNA processing factors, AtCPSF100 and AtCPSF160, but not FCA, are stably associated with FY in vivo and form a range of different-sized complexes. A hypomorphic fy allele producing a shorter protein, able to provide some FY functions but unable to interact with FCA, reduces abundance of some of the larger MW complexes. Suppressor mutants, which specifically disrupt the FY motif through which FCA interacts, also lacked these larger complexes. Our data support a model whereby FCA, perhaps after recognition of a specific RNA feature, transiently interacts with FY, an integral component of the canonical RNA 3' processing machinery, changing the interactions of the different RNA processing components. These altered interactions would appear to be a necessary step in this RNA-mediated chromatin silencing.

KW - Alleles

KW - Arabidopsis

KW - Arabidopsis Proteins

KW - Chromatin

KW - Cleavage And Polyadenylation Specificity Factor

KW - DNA Methylation

KW - Gene Expression Regulation, Plant

KW - Gene Silencing

KW - MADS Domain Proteins

KW - Mutation

KW - Nucleic Acid Conformation

KW - Protein Binding

KW - Protein Isoforms

KW - RNA, Plant

KW - RNA-Binding Proteins

KW - Transcription, Genetic

KW - mRNA Cleavage and Polyadenylation Factors

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1073/pnas.0903444106

DO - 10.1073/pnas.0903444106

M3 - Journal article

C2 - 19439664

SN - 0027-8424

VL - 106

SP - 8772

EP - 8777

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 21

ER -