Malagnac Fabienne, Bartee Lisa, Bender Judith
Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health, 615 N. Wolfe Street, Baltimore, MD 21205, USA.
EMBO J. 2002 Dec 16;21(24):6842-52. doi: 10.1093/emboj/cdf687.
Cytosine methylation is critical for correct development and genome stability in mammals and plants. In order to elucidate the factors that control genomic DNA methylation patterning, a genetic screen for mutations that disrupt methylation-correlated silencing of the endogenous gene PAI2 was conducted in Arabidopsis: This screen yielded seven loss-of-function alleles in a SET domain protein with histone H3 Lys9 methyltransferase activity, SUVH4. The mutations conferred reduced cytosine methylation on PAI2, especially in non-CG sequence contexts, but did not affect methylation on another PAI locus carrying two genes arranged as an inverted repeat. Moreover, an unmethylated PAI2 gene could be methylated de novo in the suvh4 mutant background. These results suggest that SUVH4 is involved in maintenance but not establishment of methylation at particular genomic regions. In contrast, a heterochromatin protein 1 homolog, LHP1, had no effect on PAI methylation.
胞嘧啶甲基化对于哺乳动物和植物的正常发育及基因组稳定性至关重要。为了阐明控制基因组DNA甲基化模式的因素,在拟南芥中针对破坏内源性基因PAI2甲基化相关沉默的突变进行了遗传筛选:该筛选在具有组蛋白H3赖氨酸9甲基转移酶活性的SET结构域蛋白SUVH4中产生了7个功能丧失等位基因。这些突变使PAI2上的胞嘧啶甲基化减少,尤其是在非CG序列背景下,但不影响携带两个以反向重复排列的基因的另一个PAI位点的甲基化。此外,在suvh4突变体背景下,未甲基化的PAI2基因可以从头甲基化。这些结果表明,SUVH4参与特定基因组区域甲基化的维持而非建立。相比之下,异染色质蛋白1同源物LHP1对PAI甲基化没有影响。