title: Independent chromatin binding of ARGONAUTE4 and SPT5L/KTF1 mediates transcriptional gene silencing.
abstract: Eukaryotic genomes contain significant amounts of transposons and repetitive DNA elements, which, if transcribed, can be detrimental to the organism. Expression of these elements is suppressed by establishment of repressive chromatin modifications. In Arabidopsis thaliana, they are silenced by the siRNA-mediated transcriptional gene silencing pathway where long non-coding RNAs (lncRNAs) produced by RNA Polymerase V (Pol V) guide ARGONAUTE4 (AGO4) to chromatin and attract enzymes that establish repressive chromatin modifications. It is unknown how chromatin modifying enzymes are recruited to chromatin. We show through chromatin immunoprecipitation (ChIP) that SPT5L/KTF1, a silencing factor and a homolog of SPT5 elongation factors, binds chromatin at loci subject to transcriptional silencing. Chromatin binding of SPT5L/KTF1 occurs downstream of RNA Polymerase V, but independently from the presence of 24-nt siRNA. We also show that SPT5L/KTF1 and AGO4 are recruited to chromatin in parallel and independently of each other. As shown using methylation-sensitive restriction enzymes, binding of both AGO4 and SPT5L/KTF1 is required for DNA methylation and repressive histone modifications of several loci. We propose that the coordinate binding of SPT5L and AGO4 creates a platform for direct or indirect recruitment of chromatin modifying enzymes.
authors: Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, Michigan, United States of America.
authors: M Jordan
authors: MJ
authors: Rowley
authors: Maria I
authors: MI
authors: Avrutsky
authors: Christopher J
authors: CJ
authors: Sifuentes
authors: Ligia
authors: L
authors: Pereira
authors: Andrzej T
authors: AT
authors: Wierzbicki
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000378:metabolism*; D060565:Argonaute Proteins; D002843:Chromatin / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D056426:Genetic Loci / Q000235:genetics; D008957:Models, Genetic; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: C471209:AGO4 protein, Arabidopsis; D029681:Arabidopsis Proteins; D060565:Argonaute Proteins; D002843:Chromatin; C542552:KTF1 protein, Arabidopsis; D034741:RNA, Small Interfering; D014157:Transcription Factors; C538841:RNA polymerase V, Arabidopsis; D012321:DNA-Directed RNA Polymerases