The SET Domain Proteins SUVH2 and SUVH9 Are Required for Pol V Occupancy at RNA-Directed DNA Methylation Loci (original) (raw)
Figure 6
Effect of suvh2 and suvh9 on DNA methylation and histone H3K9me2 at RdDM targets.
(A) The effect of suvh2, suvh9, and suvh2suvh9 on DNA methylation as indicated by chop-PCR. DNA methylation was determined for AtSN1, IGN5, IGN25, and solo LTR. Genomic DNA from the wild type, nrpd1, nrpe1, dms3, suvh2, suvh9, and suvh2suvh9 was cleaved by the DNA methylation-sensitive restriction enzyme HaeIII or AluI and subjected to semiquantitative PCR. The actin gene, which lacks HaeIII recognition site, was amplified as an internal control. The solo LTR locus was amplified using uncut genomic DNA as template for a control. (B) Effect of suvh2suvh9 on histone H3K9me2 was measured by CHIP assay with anti-H3K9me2 antibody. The RdDM mutants nrpe1 and dms3 were included as controls, in which the H3K9me2 levels at indicated RdDM loci were decreased. The actin gene, which lacks detectable H3K9me2, was used as a negative control.