Bestor T, Laudano A, Mattaliano R, Ingram V
Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.
J Mol Biol. 1988 Oct 20;203(4):971-83. doi: 10.1016/0022-2836(88)90122-2.
A cDNA encoding DNA (cytosine-5)-methyltransferase (DNA MeTase) of mouse cells has been cloned and sequenced. The nucleotide sequence contains an open reading frame sufficient to encode a polypeptide of 1573 amino acid residues, which is close to the apparent size of the largest species of DNA MeTase found in mouse cells. The carboxylterminal 570 amino acid residues of the inferred protein sequence shows striking similarities to bacterial type II DNA cytosine methyltransferases and appears to represent a catalytic methyltransferase domain. The amino-terminal portion of the molecule may be involved in regulating the activity of the carboxyl-terminal methyltransferase domain, since antibodies directed against a peptide sequence located within this region inhibits transmethylase activity in vitro. A 5200 base DNA MeTase-specific mRNA was found to be expressed in all mouse cell types tested, and cell lines known to have different genomic methylation patterns were found to contain DNA MeTase proteins of similar or identical sizes and de novo sequence specificities. The implications of these findings for an understanding of the mechanisms involved in the establishment and maintenance of methylation patterns are discussed.
已克隆并测序了编码小鼠细胞DNA(胞嘧啶-5)-甲基转移酶(DNA甲基转移酶)的cDNA。核苷酸序列包含一个足以编码1573个氨基酸残基多肽的开放阅读框,这与在小鼠细胞中发现的最大种类DNA甲基转移酶的表观大小相近。推测的蛋白质序列的羧基末端570个氨基酸残基与细菌II型DNA胞嘧啶甲基转移酶有显著相似性,似乎代表一个催化甲基转移酶结构域。分子的氨基末端部分可能参与调节羧基末端甲基转移酶结构域的活性,因为针对该区域内肽序列的抗体在体外可抑制甲基转移酶活性。发现一种5200个碱基的DNA甲基转移酶特异性mRNA在所有测试的小鼠细胞类型中均有表达,并且已知具有不同基因组甲基化模式的细胞系含有大小相似或相同且具有从头序列特异性的DNA甲基转移酶蛋白。讨论了这些发现对于理解甲基化模式建立和维持所涉及机制的意义。