Abstract
Nuclear RNAi provides a highly tractable system to study RNA-mediated chromatin changes and epigenetic inheritance. Recent studies have indicated that the regulation and function of nuclear RNAi-mediated heterochromatin are highly complex. Our knowledge of histone modifications and the corresponding histonemodifying enzymes involved in the system remains limited. In this study, we show that the heterochromatin mark, H3K23me3, is induced by nuclear RNAi at both exogenous and endogenous targets in C. elegans. In addition, dsRNA-induced H3K23me3 can persist for multiple generations after the dsRNA exposure has stopped. We demonstrate that the histone methyltransferase SET-32, methylates H3K23 in vitro. Both set-32 and the germline nuclear RNAi Argonaute, hrde-1, are required for nuclear RNAi-induced H3K23me3 in vivo. Our data poise H3K23me3 as an additional chromatin modification in the nuclear RNAi pathway and provides the field with a new target for uncovering the role of heterochromatin in transgenerational epigenetic silencing.
Highlights
Nuclear RNAi is an evolutionarily conserved pathway in which small RNA mediates1-4 transcriptional silencing and heterochromatin formation
We demonstrate that the histone in vitro. set-32 methyltransferase SET-32, methylates H3K23
We show that H3K23me[3] can be induced by exogenous dsRNA and is heritable for four generations
Summary
Nuclear RNAi is an evolutionarily conserved pathway in which small RNA mediates. Nuclear RNAi plays an important role in genome stability and germline development. It is a highly tractable system for the study of RNA-mediated chromatin regulation and epigenetic inheritance. C. elegans 5 provides a number of unique advantages for the study of nuclear RNAi. Genetic screens have identified numerous protein factors involved in this pathway.
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