Granlund M, Michel F, Norgren M
Department of Clinical Bacteriology, Umeå University, S-901 85 Umeå, Sweden.
J Bacteriol. 2001 Apr;183(8):2560-9. doi: 10.1128/JB.183.8.2560-2569.2001.
The present study shows that active, self-splicing group II intron GBSi1 is located downstream of the C5a-peptidase gene, scpB, in some group B streptococcus (GBS) isolates that lack insertion sequence IS1548. IS1548 was previously reported to be often present at the scpB locus in GBS isolated in association with endocarditis. Since none of 67 GBS isolates examined, 40 of which were of serotype III, harbored both IS1548 and GBSi1, these two elements are suggested to be markers for different genetic lineages in GBS serotype III. The DNA region downstream of scpB in GBS isolates harboring either GBSi1, IS1548, or none of these mobile elements was found to encode the laminin binding protein, Lmb, which shows sequence similarities to a family of streptococcal adhesins. IS1548 is inserted 9 bp upstream of the putative promoter for lmb, while the insertion site for GBSi1 is located 88 bp further upstream. Sequences highly similar to GBSi1 exist also in Streptococcus pneumoniae. An inverted repeat sequence, with features typical of transcription terminators, was identified immediately upstream of the insertion site for the group II intron both in the GBS and S. pneumoniae sequences. This motif is suggested to constitute a target for the GBS intron as well as for rather closely related introns in Bacillus halodurans, Pseudomonas alcaligenes, and Pseudomonas putida. When transcripts containing the GBSi1 intron were incubated at high concentrations of ammonium and magnesium, a major product with the expected length and sequence for the ligated exons was generated. Unlike, however, all members of group II investigated so far, the excised intron was in linear, rather than in a branched (lariat), form.
本研究表明,活性自剪接II组内含子GBSi1位于某些缺乏插入序列IS1548的B族链球菌(GBS)分离株中C5a肽酶基因scpB的下游。此前报道,IS1548常出现在与心内膜炎相关的GBS分离株的scpB位点。由于所检测的67株GBS分离株中无一同时携带IS1548和GBSi1,其中40株为III型血清型,因此这两个元件被认为是GBS III型血清型中不同遗传谱系的标志物。在携带GBSi1、IS1548或均不携带这些移动元件的GBS分离株中,scpB下游的DNA区域被发现编码层粘连蛋白结合蛋白Lmb,其与链球菌粘附素家族具有序列相似性。IS1548插入到lmb推定启动子上游9 bp处,而GBSi1的插入位点则位于更上游88 bp处。肺炎链球菌中也存在与GBSi1高度相似的序列。在GBS和肺炎链球菌序列中,在II组内含子插入位点的紧邻上游均鉴定出一个具有转录终止子典型特征的反向重复序列。该基序被认为是GBS内含子以及嗜碱芽孢杆菌、产碱假单胞菌和恶臭假单胞菌中亲缘关系较近的内含子的靶点。当含有GBSi1内含子的转录本在高浓度铵和镁条件下孵育时,产生了具有预期长度和连接外显子序列的主要产物。然而,与迄今研究的所有II组成员不同,切除的内含子呈线性而非分支(套索)形式。