Clr4 Ubiquitination and Non-coding RNA Mediate Transcriptional Silencing of Heterochromatin Via Swi6 Phase Separation

Overview

Journal Nat Commun

Specialty Biology

Date 2024 Oct 31

PMID 39477922

Authors

Hyun-Soo Kim

Benjamin Roche

Sonali Bhattacharjee

Leila Todeschini

An-Yun Chang

Christopher Hammell

Andre Verdel

Robert A Martienssen

Affiliations

Soon will be listed here.

Abstract

Transcriptional silencing by RNAi paradoxically relies on transcription, but how the transition from transcription to silencing is achieved has remained unclear. The Cryptic Loci Regulator complex (CLRC) in Schizosaccharomyces pombe is a cullin-ring E3 ligase required for silencing that is recruited by RNAi. We found that the E2 ubiquitin conjugating enzyme Ubc4 interacts with CLRC and mono-ubiquitinates the histone H3K9 methyltransferase Clr4, promoting the transition from co-transcriptional gene silencing (H3K9me2) to transcriptional gene silencing (H3K9me3). Ubiquitination of Clr4 occurs in an intrinsically disordered region (Clr4), which undergoes liquid droplet formation in vitro, along with Swi6 the effector of transcriptional gene silencing. Our data suggests that phase separation is exquisitely sensitive to non-coding RNA (ncRNA) which promotes self-association of Clr4, chromatin association, and di-, but not tri- methylation instead. Ubc4-CLRC also targets the transcriptional co-activator Bdf2, down-regulating centromeric transcription and small RNA (sRNA) production. The deubiquitinase Ubp3 counteracts both activities.

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