DNA methylation promotes transcription: DNA methylation generally represses transcription, but in some instances, it has also been implicated in transcription activation. Harris et al. identified a protein complex in Arabidopsis that is recruited to chromatin by DNA methylation. This complex specifically activated the transcription of genes that are already mildly transcribed but had no effect on transcriptionally silent genes such as transposable elements. The complex thereby counteracts the repression effect caused by transposon insertion in neighboring genes while leaving transposons silent. Thus, by balancing both repressive and activating transcriptional effects, DNA methylation can act to fine-tune gene expression. Science , this issue p. 1182

Significance: Plant DNA methylation, which occurs in three sequence contexts (CG, CHG, and CHH, where H refers to A, T, or C), is established and maintained by different mechanisms. In this study, we present genome-wide chromatin accessibility profiles of Arabidopsis mutants that are deficient in CG, CHG, and/or CHH methylation. Through a combination of DNA methylation, chromatin accessibility, and higher-order chromosome conformation profiling of these mutants, we uncover links between DNA methylation, chromatin accessibility, and 3D genome architecture. These results reveal the interplay between CG and non-CG methylation in heterochromatin maintenance and suggest that DNA methylation can directly impact chromatin structure.

DNA methylation functions in gene silencing and the maintenance of genome integrity. In plants, non-CG DNA methylation is linked through a self-reinforcing loop with histone 3 lysine 9 dimethylation (H3K9me2). The plant-specific SUPPRESSOR OF VARIEGATION 3–9 HOMOLOG (SUVH) family H3K9 methyltransferases (MTases) bind to DNA methylation marks and catalyze H3K9 methylation. Here, we analyzed the structure and function of Arabidopsis thaliana SUVH6 to understand how this class of enzyme maintains methylation patterns in the genome. We reveal that SUVH6 has a distinct 5-methyl-dC (5mC) base-flipping mechanism involving a thumb loop element. Autoinhibition of H3 substrate entry is regulated by a SET domain loop, and a conformational transition in the post-SET domain upon cofactor binding may control catalysis. In vitro DNA binding and in vivo ChIP-seq data reveal that the different SUVH family H3K9 MTases have distinct DNA binding preferences, targeting H3K9 methylation to sites with different methylated DNA sequences, explaining the context biased non-CG DNA methylation in plants. Significance Plant SUVH family H3K9 methyltransferases play a key role in connecting the two epigenetic silencing marks, DNA methylation and H3K9me2. However, the regulation of SUVH protein activities and their precise role in the regulation of DNA methylation remains unclear. In this research, we performed a comprehensive investigation into the structure, biochemistry, and in vivo targeting characteristics of SUVH histone methyltransferases. For binding methylated DNA, we reveal that the SUVH family proteins possess a unique thumb loop-dependent base-flipping mechanism. For methyltransferase function, we reveal that SUVH6 is regulated by a dynamic autoinhibitory domain. Finally, our in vitro DNA-binding assays combined with ChIP-seq data uncover mechanisms to help explain context-biased non-CG DNA methylation in plants.

Ready to dice: In the biogenesis of small RNAs, the Dicer family endonucleases act as a molecular ruler to cut the substrate RNA into defined lengths. Wang et al . report a structure of the dicing-competent state of plant DICER LIKE PROTEIN 3 (DCL3) in complex with a pre–small interfering RNA (pre-siRNA). Toward one end of the pre-siRNA, DCL3 uses a positively charged pocket and an aromatic cap to specifically recognize the 5′-phosphorylated adenosine of the guide strand and the 3′ overhang of the complementary strand, respectively. On the other end, the paired ribonuclease III domains of DCL3 cut both strands of the RNA, determining the precise length of the product small RNA. —DJ