Johnson Lianna M, Bostick Magnolia, Zhang Xiaoyu, Kraft Edward, Henderson Ian, Callis Judy, Jacobsen Steven E
Life Science Core Curriculum, University of California, Los Angeles, Los Angeles, California 90095, USA.
Curr Biol. 2007 Feb 20;17(4):379-84. doi: 10.1016/j.cub.2007.01.009. Epub 2007 Jan 18.
Epigenetic gene silencing suppresses transposon activity and is critical for normal development . Two common epigenetic gene-silencing marks are DNA methylation and histone H3 lysine 9 dimethylation (H3K9me2). In Arabidopsis thaliana, H3K9me2, catalyzed by the methyltransferase KRYPTONITE (KYP/SUVH4), is required for maintenance of DNA methylation outside of the standard CG sequence context. Additionally, loss of DNA methylation in the met1 mutant correlates with a loss of H3K9me2. Here we show that KYP-dependent H3K9me2 is found at non-CG methylation sites in addition to those rich in CG methylation. Furthermore, we show that the SRA domain of KYP binds directly to methylated DNA, and SRA domains with missense mutations found in loss-of-function kyp mutants have reduced binding to methylated DNA in vitro. These data suggest that DNA methylation is required for the recruitment or activity of KYP and suggest a self-reinforcing loop between histone and DNA methylation. Lastly, we found that SRA domains from two Arabidopsis SRA-RING proteins also bind methylated DNA and that the SRA domains from KYP and SRA-RING proteins prefer methylcytosines in different sequence contexts. Hence, unlike the methyl-binding domain (MBD), which binds only methylated-CpG sequences, the SRA domain is a versatile new methyl-DNA-binding motif.
表观遗传基因沉默抑制转座子活性,对正常发育至关重要。两种常见的表观遗传基因沉默标记是DNA甲基化和组蛋白H3赖氨酸9二甲基化(H3K9me2)。在拟南芥中,由甲基转移酶KRYPTONITE(KYP/SUVH4)催化的H3K9me2是维持标准CG序列上下文之外的DNA甲基化所必需的。此外,met1突变体中DNA甲基化的缺失与H3K9me2的缺失相关。在这里我们表明,除了富含CG甲基化的位点外,KYP依赖的H3K9me2还存在于非CG甲基化位点。此外,我们表明KYP的SRA结构域直接与甲基化DNA结合,并且在功能缺失的kyp突变体中发现的具有错义突变的SRA结构域在体外与甲基化DNA的结合减少。这些数据表明DNA甲基化是KYP募集或活性所必需的,并提示组蛋白和DNA甲基化之间存在自我强化循环。最后,我们发现来自两种拟南芥SRA-RING蛋白的SRA结构域也结合甲基化DNA,并且来自KYP和SRA-RING蛋白的SRA结构域在不同的序列上下文中更喜欢甲基胞嘧啶。因此,与仅结合甲基化-CpG序列的甲基结合结构域(MBD)不同,SRA结构域是一种通用的新型甲基-DNA结合基序。