Peptide mapping of the murine DNA methyltransferase reveals a major phosphorylation site and the start of translation
Peptide mapping of the murine DNA methyltransferase reveals a major phosphorylation site and the start of translation
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DOI：
10.1074/jbc.272.28.17851
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发表时间：
1997-07-11
影响因子：
4.8
通讯作者：
Reich, NO
中科院分区：
文献类型：
--
作者：
Glickman, JF;Pavlovich, JG;Reich, NO
小鼠DNA甲基转移酶催化甲基从s -腺苷蛋氨酸转移到d（CpG）二核苷酸内的胞嘧啶。该酶是正常胚胎发育所必需的，并涉及许多重要的过程，包括基因表达和癌症的控制。对从小鼠红白血病细胞纯化的DNA甲基转移酶进行了代谢标记和高压液相色谱-电喷雾电离-质谱（HPLC-ESI-MS）。丝氨酸514被确定为一个主要的磷酸化位点，位于酶靶向复制中心所需的区域，这些结果提供了DNA甲基化调控的潜在机制。HPLC-ESI-MS多肽定位数据表明，纯化的小鼠DNA甲基转移酶蛋白含有最近修订的5'基因序列预测的n端区域（Yoder， J. A., Yen， R.-W.）。C., Vertino， P. M., Bestor， T. H., and Baylin， S. B. (1996) J.生物学，化学，271,31092-31097)，证据表明翻译开始于第一个预测的蛋氨酸，没有替代的翻译起始位点。我们的肽图谱结果提供了更详细的DNA甲基转移酶的结构表征，将有助于未来的结构/功能研究。
The murine DNA methyltransferase catalyzes the transfer of methyl groups from S-adenosylmethionine to cytosines within d(CpG) dinucleotides. The enzyme is necessary for normal embryonic development and is implicated in a number of important processes, including the control of gene expression and cancer, Metabolic labeling and high pressure liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) were performed on DNA methyltransferase purified from murine erythroleukemia cells, Serine 514 was identified as a major phosphorylation site that lies in a domain required for targeting of the enzyme to the replication foci, These results present a potential mechanism for the regulation of DNA methylation.HPLC-ESI-MS peptide mapping data demonstrated that the purified murine DNA methyltransferase protein contains the N-terminal regions predicted by the recently revised 5' gene sequences (Yoder, J. A., Yen, R.-W. C., Vertino, P. M., Bestor, T. H., and Baylin, S. B. (1996) J. Biol, Chem, 271, 31092-31097), The evidence suggests a start of translation at the first predicted methionine, with no alternate translational start sites, Our peptide mapping results provide a more detailed structural characterization of the DNA methyltransferase that will facilitate future structure/function studies.