Scavetta R D, Thomas C B, Walsh M A, Szegedi S, Joachimiak A, Gumport R I, Churchill M E
Department of Pharmacology, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, CO 80262, USA.
Nucleic Acids Res. 2000 Oct 15;28(20):3950-61. doi: 10.1093/nar/28.20.3950.
DNA methylation is important in cellular, developmental and disease processes, as well as in bacterial restriction-modification systems. Methylation of DNA at the amino groups of cytosine and adenine is a common mode of protection against restriction endonucleases afforded by the bacterial methyltransferases. The first structure of an N:6-adenine methyltransferase belonging to the beta class of bacterial methyltransferases is described here. The structure of M. RSR:I from Rhodobacter sphaeroides, which methylates the second adenine of the GAATTC sequence, was determined to 1.75 A resolution using X-ray crystallography. Like other methyltransferases, the enzyme contains the methylase fold and has well-defined substrate binding pockets. The catalytic core most closely resembles the PVU:II methyltransferase, a cytosine amino methyltransferase of the same beta group. The larger nucleotide binding pocket observed in M. RSR:I is expected because it methylates adenine. However, the most striking difference between the RSR:I methyltransferase and the other bacterial enzymes is the structure of the putative DNA target recognition domain, which is formed in part by two helices on an extended arm of the protein on the face of the enzyme opposite the active site. This observation suggests that a dramatic conformational change or oligomerization may take place during DNA binding and methylation.
DNA甲基化在细胞、发育和疾病过程以及细菌限制修饰系统中都很重要。胞嘧啶和腺嘌呤氨基上的DNA甲基化是细菌甲基转移酶提供的一种常见的抗限制性内切酶保护模式。本文描述了属于细菌甲基转移酶β类的N:6-腺嘌呤甲基转移酶的首个结构。利用X射线晶体学确定了来自球形红细菌的M. RSR:I的结构,其分辨率为1.75 Å,该酶可将GAATTC序列的第二个腺嘌呤甲基化。与其他甲基转移酶一样,该酶含有甲基化酶折叠结构,并有明确的底物结合口袋。催化核心与PVU:II甲基转移酶最为相似,PVU:II是同一β类的胞嘧啶氨基甲基转移酶。在M. RSR:I中观察到的较大的核苷酸结合口袋是预期的,因为它甲基化腺嘌呤。然而,RSR:I甲基转移酶与其他细菌酶之间最显著的差异在于假定的DNA靶标识别结构域的结构,该结构部分由位于酶活性位点对面的蛋白质延伸臂上的两个螺旋形成。这一观察结果表明,在DNA结合和甲基化过程中可能会发生显著的构象变化或寡聚化。