title: The role of the Arabidopsis Exosome in siRNA-independent silencing of heterochromatic loci.
abstract: The exosome functions throughout eukaryotic RNA metabolism and has a prominent role in gene silencing in yeast. In Arabidopsis, exosome regulates expression of a "hidden" transcriptome layer from centromeric, pericentromeric, and other heterochromatic loci that are also controlled by small (sm)RNA-based de novo DNA methylation (RdDM). However, the relationship between exosome and smRNAs in gene silencing in Arabidopsis remains unexplored. To investigate whether exosome interacts with RdDM, we profiled Arabidopsis smRNAs by deep sequencing in exosome and RdDM mutants and also analyzed RdDM-controlled loci. We found that exosome loss had a very minor effect on global smRNA populations, suggesting that, in contrast to fission yeast, in Arabidopsis the exosome does not control the spurious entry of RNAs into smRNA pathways. Exosome defects resulted in decreased histone H3K9 dimethylation at RdDM-controlled loci, without affecting smRNAs or DNA methylation. Exosome also exhibits a strong genetic interaction with RNA Pol V, but not Pol IV, and physically associates with transcripts produced from the scaffold RNAs generating region. We also show that two Arabidopsis rrp6 homologues act in gene silencing. Our data suggest that Arabidopsis exosome may act in parallel with RdDM in gene silencing, by epigenetic effects on chromatin structure, not through siRNAs or DNA methylation.
authors.affiliation: School of Biological Sciences, University of Missouri-Kansas City, Kansas City, Missouri, United States of America.
authors.forename: Jun-Hye
authors.initials: JH
authors.lastname: Shin
authors.forename: Hsiao-Lin V
authors.initials: HL
authors.lastname: Wang
authors.forename: Jinwon
authors.initials: J
authors.lastname: Lee
authors.forename: Brandon L
authors.initials: BL
authors.lastname: Dinwiddie
authors.forename: Dmitry A
authors.initials: DA
authors.lastname: Belostotsky
authors.forename: Julia A
authors.initials: JA
authors.lastname: Chekanova
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation; D055354:Exosomes / Q000235:genetics / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D006570:Heterochromatin / Q000235:genetics* / Q000378:metabolism; D059014:High-Throughput Nucleotide Sequencing; D012313:RNA / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D006570:Heterochromatin; D034741:RNA, Small Interfering; D012313:RNA
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