一种在体外衰老系统中研究DNA甲基转移酶表达的方法。
A method to study the expression of DNA methyltransferases in aging systems in vitro.
作者信息
Berletch Joel B, Phipps Sharla M O, Walthall Sabrina L, Andrews Lucy G, Tollefsbol Trygve O
机构信息
Department of Biology, University of Alabama at Birmingham, Birmingham, AL, USA.
出版信息
Methods Mol Biol. 2007;371:81-7. doi: 10.1007/978-1-59745-361-5_7.
The methylation of CpG dinucleotides located in key protein binding sites within gene regulatory regions often leads to gene silencing. A mechanism of aging is proposed whereby an accumulation of methylation at gene regulatory sites contributes to cellular senescence. DNA methyltransferases (DNMTs) are enzymes that catalyze the transfer of a methyl moiety from S-adenosyl-L-methionine (SAM) to the cytosine of a CpG dinucleotide and are responsible for establishing and maintaining methylation patterns in the genome. It is important to study not only transcription of the DNMTs, but also their protein expression because studies illustrate that it is possible for the enzymes to undergo posttranslational physical changes in response to stimulation even though gene transcription remains unchanged. Here, we discuss an in vitro method to study protein expression of DNMTs in aging systems.
位于基因调控区域内关键蛋白质结合位点的CpG二核苷酸甲基化通常会导致基因沉默。有人提出了一种衰老机制，即基因调控位点甲基化的积累导致细胞衰老。DNA甲基转移酶（DNMTs）是一类酶，可催化甲基基团从S-腺苷-L-甲硫氨酸（SAM）转移至CpG二核苷酸的胞嘧啶上，并负责在基因组中建立和维持甲基化模式。不仅要研究DNMTs的转录，还要研究它们的蛋白质表达，这一点很重要，因为研究表明，即使基因转录保持不变，这些酶也可能会因刺激而发生翻译后物理变化。在此，我们讨论一种在衰老系统中研究DNMTs蛋白质表达的体外方法。
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