Project description:BACKGROUND: Polycomb group complexes PRC1 and PRC2 repress gene expression at the chromatin level in eukaryotes. The classic recruitment model of Polycomb group complexes in which PRC2-mediated H3K27 trimethylation recruits PRC1 for H2A monoubiquitination was recently challenged by data showing that PRC1 activity can also recruit PRC2. However, the prevalence of these two mechanisms is unknown, especially in plants as H2AK121ub marks were examined at only a handful of Polycomb group targets. RESULTS: By using genome-wide analyses, we show that H2AK121ub marks are surprisingly widespread in Arabidopsis thaliana, often co-localizing with H3K27me3 but also occupying a set of transcriptionally active genes devoid of H3K27me3. Furthermore, by profiling H2AK121ub and H3K27me3 marks in atbmi1a/b/c, clf/swn, and lhp1 mutants we found that PRC2 activity is not required for H2AK121ub marking at most genes. In contrast, loss of AtBMI1 function impacts the incorporation of H3K27me3 marks at most Polycomb group targets. CONCLUSIONS: Our findings show the relationship between H2AK121ub and H3K27me3 marks across the A. thaliana genome and unveil that ubiquitination by PRC1 is largely independent of PRC2 activity in plants, while the inverse is true for H3K27 trimethylation.
Project description:BACKGROUND: Polycomb group complexes PRC1 and PRC2 repress gene expression at the chromatin level in eukaryotes. The classic recruitment model of Polycomb group complexes in which PRC2-mediated H3K27 trimethylation recruits PRC1 for H2A monoubiquitination was recently challenged by data showing that PRC1 activity can also recruit PRC2. However, the prevalence of these two mechanisms is unknown, especially in plants as H2AK121ub marks were examined at only a handful of Polycomb group targets. RESULTS: By using genome-wide analyses, we show that H2AK121ub marks are surprisingly widespread in Arabidopsis thaliana, often co-localizing with H3K27me3 but also occupying a set of transcriptionally active genes devoid of H3K27me3. Furthermore, by profiling H2AK121ub and H3K27me3 marks in atbmi1a/b/c, clf/swn, and lhp1 mutants we found that PRC2 activity is not required for H2AK121ub marking at most genes. In contrast, loss of AtBMI1 function impacts the incorporation of H3K27me3 marks at most Polycomb group targets. CONCLUSIONS: Our findings show the relationship between H2AK121ub and H3K27me3 marks across the A. thaliana genome and unveil that ubiquitination by PRC1 is largely independent of PRC2 activity in plants, while the inverse is true for H3K27 trimethylation.
Project description:Histone H2A monoubiquitination (H2Aub1) function as a conserved post-translational modification in eukaryotes to maintain gene repression and ensure cellular identity. Arabidopsis H2Aub1 is catalyzed by polycomb repressive complex 1 (PRC1) contributed by AtRING1s and AtBMI1s. Because PRC1 components lack known DNA binding domains, how H2Aub1 is established at the specific genomic sites remains a topic of debate. Here we show that the cohesin subunits AtSYN4 and AtSCC3 interact with each other and AtSCC3 binds to AtBMI1s. We find that H2Aub1 levels are significantly reduced in the atsyn4 mutant or AtSCC3 RNAi knockdown plants. ChIP-seq analysis indicated that AtSYN4 and AtSCC3 tend to associate with H2Aub1 in the genome regions of transcription activation independent on H3K27me3. Finally, we show that AtSYN4 binds directly to the G-box element and targets H2Aub1to these sites. Our study thus revealed a mechanism for cohesin-mediated recruitment of AtBMI1s to specific loci to mediate H2Aub1.
Project description:Histone H2A monoubiquitination (H2Aub1) is a ubiquitous and conserved post-translational modification, playing a non-degradative role in multiple progresses including transcription regulation. In Arabidopsis thaliana, H2Aub1 is mediated by AtRING1s and AtBMI1s, catalytic subunits of Polycomb repressive complex 1 (PRC1). Despite PRC2-associated repressive role of H2Aub1, the mechanism regarding how H2Aub1 distributed on non-H3K27me3 region need to be explained. In this article, we show that the two cohesin subunits AtSCC3 and AtSYN4 directly interact with AtBMI1A/B/C, and theH2Aub1 levels are significantly reduced in atsyn4orAtSCC3RNAi plants. By ChIP-seq analysis, we found that AtSYN4 and AtSCC3 co-localize with H2Aub1 in genome, and are related to H3K27me3-independent transcription activation. Further, we provide evidences to show that AtSYN4 directly bind to G-box element and regulate H2Aub1 modification in G-box containing loci. These findings reveal that AtSCC3 and AtSYN4 are directly and functionally associated with AtBMI1s. Our study uncovers the mechanism of H2Aub1 targeting modification.