Brevnov M G, Kubareva E A, Romanova E A, Volkov E M, Karyagina A S, Nikolskaya I I, Gromova E S
A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia.
Gene. 1995 May 19;157(1-2):149-52. doi: 10.1016/0378-1119(94)00738-e.
The interaction of the MvaI and SsoII DNA methyltransferases (MTases; M.MVaI and M.SsoII, respectively) with a set of synthetic DNA duplexes, containing a M.MvaI and M.SsoII recognition site (CCWGG), was investigated. In these DNA duplexes dA or dT of the recognition site was replaced by nucleoside analogs with modified sugar moieties and heterocyclic bases (2'-deoxy-2'-fluorouridine (flU), 1-(beta-D-2'-deoxy-threo-pentofuranosyl)thymine (xT), 1-(beta-D-3'-deoxy-threo-pentofuranosyl)uracil (tU)), or by 1,3-propanediol (Prd). A new approach for monitoring methylation of each strand of DNA duplexes by MTases was developed. It allowed the determination of the influence of the modification in one DNA strand on the methylation of the other. In most cases, for both M.MvaI and M.SsoII, sugar analog-containing duplexes showed inhibition of methylation of only the modified strand. Prd-containing DNA duplexes were not substrates for M.MvaI. M.SsoII did not methylate DNA duplexes in which the dT residue was replaced by Prd.
研究了MvaI和SsoII DNA甲基转移酶（MTases；分别为M.MVaI和M.SsoII）与一组合成DNA双链体的相互作用，这些双链体含有M.MvaI和M.SsoII识别位点（CCWGG）。在这些DNA双链体中，识别位点的dA或dT被具有修饰糖部分和杂环碱基的核苷类似物（2'-脱氧-2'-氟尿苷（flU）、1-（β-D-2'-脱氧-苏式-戊呋喃糖基）胸腺嘧啶（xT）、1-（β-D-3'-脱氧-苏式-戊呋喃糖基）尿嘧啶（tU））或1,3-丙二醇（Prd）取代。开发了一种监测MTases对DNA双链体每条链甲基化的新方法。它能够确定一条DNA链上的修饰对另一条链甲基化的影响。在大多数情况下，对于M.MvaI和M.SsoII而言，含糖类似物的双链体仅显示对修饰链甲基化的抑制作用。含Prd的DNA双链体不是M.MvaI的底物。M.SsoII不会使dT残基被Prd取代的DNA双链体甲基化。
