一种在衰老系统中体外检测DNA甲基转移酶I基因转录的方法。
A method to detect DNA methyltransferase I gene transcription in vitro in aging systems.
作者信息
Berletch Joel B, 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;371:73-80. doi: 10.1007/978-1-59745-361-5_6.
Epigenetic alterations of DNA play key roles in determining gene structure and expression. Methylation of the 5-position of cytosine is thought to be the most common modification of the genome in mammals. Studies have generally shown that hypermethylation in gene regulatory regions is associated with inactivation and reduced transcription and that alteration in established methylation patterns during development can affect embryonic viability. Changes in methylation have also been associated with aging and cellular senescence as well as tumorogenesis. DNA methyltransferase 1 (DNMT1) is thought to play an important role in maintaining already established methylation patterns during DNA replication and catalyzes the transfer of a methyl moiety from S-adenosyl-L-methionine (SAM) to the 5-position of cytosines in the CpG dinucleotide. Several studies illustrate changes in activity and transcription of DNMT1 during aging and here we show a comprehensive method of detection of DNMT1 mRNA transcription from senescing cells in culture.
DNA的表观遗传改变在决定基因结构和表达方面起着关键作用。胞嘧啶5位的甲基化被认为是哺乳动物基因组中最常见的修饰。研究普遍表明，基因调控区域的高甲基化与基因失活和转录减少相关，并且发育过程中既定甲基化模式的改变会影响胚胎活力。甲基化的变化还与衰老、细胞衰老以及肿瘤发生有关。DNA甲基转移酶1（DNMT1）被认为在DNA复制过程中维持已建立的甲基化模式方面发挥重要作用，并催化甲基基团从S-腺苷-L-甲硫氨酸（SAM）转移至CpG二核苷酸中胞嘧啶的5位。多项研究阐明了衰老过程中DNMT1活性和转录的变化，在此我们展示了一种检测培养中衰老细胞DNMT1 mRNA转录的综合方法。
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