Ko Minsu, Choi Hyang, Park Chankyu
National Creative Research Initiative Center for Behavioral Genetics, Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Yusong-Ku, Taejon 305-701, Republic of Korea.
J Bacteriol. 2002 Jul;184(14):3917-22. doi: 10.1128/JB.184.14.3917-3922.2002.
Self-splicing introns are rarely found in bacteria and bacteriophages. They are classified into group I and II according to their structural features and splicing mechanisms. While the group I introns are occasionally found in protein-coding regions of phage genomes and in several tRNA genes of cyanobacteria and proteobacteria, they had not been found in protein-coding regions of bacterial genomes. Here we report a group I intron in the recA gene of Bacillus anthracis which was initially found by DNA sequencing as an intervening sequence (IVS). By using reverse transcriptase PCR, the IVS was shown to be removable from the recA precursor mRNA for RecA that was being translated in E. coli. The splicing was visualized in vitro with labeled free GTP, indicating that it is a group I intron, which is also implied by its predicted secondary structure. The RecA protein of B. anthracis expressed in E. coli was functional in its ability to complement a recA defect. When recA-negative E. coli cells were irradiated with UV, the Bacillus RecA reduced the UV susceptibility of the recA mutant, regardless of the presence of intron.
自我剪接内含子在细菌和噬菌体中很少见。根据其结构特征和剪接机制,它们被分为I类和II类。虽然I类内含子偶尔会出现在噬菌体基因组的蛋白质编码区以及蓝细菌和变形菌的几个tRNA基因中,但尚未在细菌基因组的蛋白质编码区中发现。在此,我们报告了炭疽芽孢杆菌recA基因中的一个I类内含子,它最初是通过DNA测序作为一个间隔序列(IVS)被发现的。通过使用逆转录酶PCR,该IVS被证明可从在大肠杆菌中正在翻译的RecA的recA前体mRNA中去除。用标记的游离GTP在体外观察到了剪接,表明它是一个I类内含子,其预测的二级结构也暗示了这一点。在大肠杆菌中表达的炭疽芽孢杆菌RecA蛋白在互补recA缺陷的能力方面是有功能的。当recA阴性的大肠杆菌细胞受到紫外线照射时,并无论内含子是否存在,芽孢杆菌RecA都降低了recA突变体对紫外线的敏感性。