Lee K F, Shaw P C, Picone S J, Wilson G G, Lunnen K D
Department of Biochemistry, The Chinese University of Hong Kong, Shatin, NT.
Biol Chem. 1998 Apr-May;379(4-5):437-41. doi: 10.1515/bchm.1998.379.4-5.437.
The genes coding for the EcoHK31I and EaeI restriction-modification (R-M) systems from Escherichia coli strain HK31 and Enterobacter aerogenes, respectively, have been cloned and sequenced. Both ENases recognize and cleave Y/GGCCR leaving 4 nucleotide 5'-protruding ends, while the MTases modify the internal cytosine. The systems were isolated on a 2.3kb AseI fragment for EcoHK31I, and a 4.6 kb HindIII fragment for EaeI. The R and M genes of both systems converge and overlap by 14 nucleotides. Previously, we found that M.EcoHK31I consisted of two subunits, (alpha and beta), with the beta subunit being translated from an alternative open reading frame within the gene encoding the alpha subunit. Sequence comparison between the EcoHK31I and EaeI systems reveals striking similarity. The eaeIM gene also encodes alpha and beta polypeptides of 309 and 176 amino acids which share 96% and 97% identity, respectively, with those of ecoHK31IM. ecoHK31IR and eaeIR encode proteins of 318 and 315 aa, respectively, which share 92% identity but are otherwise unique in the GenBank database. The EaeI and the EcoHK31I R-M systems were found to be flanked by genes coding for integrases. It is possible that these integrases have facilitated the transfer of this system among different bacterial species.
分别来自大肠杆菌HK31菌株和产气肠杆菌的编码EcoHK31I和EaeI限制修饰(R-M)系统的基因已被克隆和测序。两种核酸内切酶都识别并切割Y/GGCCR,产生4个核苷酸的5'突出末端,而甲基转移酶则修饰内部的胞嘧啶。EcoHK31I系统是在一个2.3kb的AseI片段上分离得到的,EaeI系统是在一个4.6kb的HindIII片段上分离得到的。这两个系统的R基因和M基因相互汇聚并重叠14个核苷酸。此前,我们发现M.EcoHK31I由两个亚基(α和β)组成,β亚基是从编码α亚基的基因内的一个替代开放阅读框翻译而来的。EcoHK31I和EaeI系统之间的序列比较显示出惊人的相似性。eaeIM基因也编码309和176个氨基酸的α和β多肽,它们与ecoHK31IM的α和β多肽分别具有96%和97%的同一性。ecoHK31IR和eaeIR分别编码318和315个氨基酸的蛋白质,它们具有92%的同一性,但在GenBank数据库中其他方面是独特的。发现EaeI和EcoHK31I R-M系统两侧是编码整合酶的基因。这些整合酶有可能促进了该系统在不同细菌物种之间的转移。