Kersulyte D, Akopyants N S, Clifton S W, Roe B A, Berg D E
Departments of Molecular Microbiology and Genetics, Campus Box 8230, Washington University School of Medicine, St. Louis, MO 63110, USA.
Gene. 1998 Nov 26;223(1-2):175-86. doi: 10.1016/s0378-1119(98)00164-4.
IS605, an insertion sequence (IS) that is unusual in containing homologs of genes for the single putative transposases of two other unrelated IS elements (IS200 and IS1341), was found in nearly one-third of a set of 238 independent isolates of the gastric pathogen Helicobacter pylori. Hybridization and PCR tests indicated that any strain carrying one of these ORFs also carried the other, which implies that both ORFs are in the same unit of transposition. The IS605 ends and target sites for insertion were identified by sequencing eight preexisting insertions in strain NCTC11638, corresponding empty sites in other strains, and new transpositions in E. coli of an IS605 derivative marked with a selectable chloramphenicol-resistance gene. These tests showed that IS605 is also unusual in: (1) having unique, not inverted repeat, ends; (2) not duplicating (or deleting) target sequences during transposition; and (3) inserting with its left (IS200-homolog) end next to 5'-TTTAA or 5'-TTTAAC. IS605 was implicated in at least two genome rearrangements in strain NCTC11638. A second member of the IS605 family, called IS606 (25% amino acid identity to IS605 in inferred proteins) was found in one-third of 38 H. pylori strains tested, many of which did not carry IS605. The features of these two chimaeric IS elements are discussed in terms of possible transposition mechanisms, IS element evolution, and effects of IS elements on genome organization and evolution in the microbes that they inhabit.
IS605是一种插入序列(IS),其不同寻常之处在于它包含另外两个不相关的IS元件(IS200和IS1341)的单个假定转座酶基因的同源物,在一组238株独立的胃病原体幽门螺杆菌分离株中,近三分之一都发现了该序列。杂交和PCR检测表明,携带这些开放阅读框(ORF)之一的任何菌株也携带另一个,这意味着两个ORF都在同一个转座单位中。通过对菌株NCTC11638中八个预先存在的插入序列、其他菌株中的相应空位以及用可选择的氯霉素抗性基因标记的IS605衍生物在大肠杆菌中的新转座进行测序,确定了IS605的末端和插入靶点。这些检测还表明,IS605在以下方面也很不寻常:(1)具有独特的末端,而非反向重复末端;(2)在转座过程中不复制(或删除)靶点序列;(3)以其左侧(与IS200同源)末端紧邻5'-TTTAA或5'-TTTAAC的方式插入。IS605与菌株NCTC11638中的至少两次基因组重排有关。在38株测试的幽门螺杆菌菌株中,三分之一发现了IS605家族的第二个成员,称为IS606(在推断的蛋白质中与IS605有25%的氨基酸同一性),其中许多菌株不携带IS605。本文从可能的转座机制、IS元件进化以及IS元件对其所栖息微生物的基因组组织和进化的影响等方面讨论了这两种嵌合IS元件的特征。