Ctf4-related protein recruits LHP1-PRC2 to maintain H3K27me3 levels in dividing cells in <i>Arabidopsis thaliana</i>

Abstract

Polycomb Repressive Complex (PRC) 2 catalyzes the H3K27me3 modification that warrants inheritance of a repressive chromatin structure during cell division, thereby assuring stable target gene repression in differentiated cells. It is still under investigation how H3K27me3 is passed on from maternal to filial strands during DNA replication; however, cell division can reinforce H3K27me3 coverage at target regions. To identify novel factors involved in the Polycomb pathway in plants, we performed a forward genetic screen for enhancers of the <i>like heterochromatin protein 1</i> (<i>lhp1</i>) mutant, which shows relatively mild phenotypic alterations compared with other plant PRC mutants. We mapped <i>enhancer of lhp1</i> (<i>eol) 1</i> to a gene related to yeast <i>Chromosome transmission fidelity 4</i> (<i>Ctf4</i>) based on phylogenetic analysis, structural similarities, physical interaction with the CMG helicase component SLD5, and an expression pattern confined to actively dividing cells. A combination of <i>eol1</i> with the <i>curly leaf</i> (<i>clf</i>) allele, carrying a mutation in the catalytic core of PRC2, strongly enhanced the <i>clf</i> phenotype; furthermore, H3K27me3 coverage at target genes was strongly reduced in <i>eol1 clf</i> double mutants compared with <i>clf</i> single mutants. EOL1 physically interacted with CLF, its partially redundant paralog SWINGER (SWN), and LHP1. We propose that EOL1 interacts with LHP1-PRC2 complexes during replication and thereby participates in maintaining the H3K27me3 mark at target genes.