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 -