Huisman O, Errada P R, Signon L, Kleckner N
Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
EMBO J. 1989 Jul;8(7):2101-9. doi: 10.1002/j.1460-2075.1989.tb03619.x.
We present the genetic analysis of a large number of mutations in the outside end of insertion sequence IS10. (i) The terminal inverted repeat sequence is probably the primary site of transposase binding. Mutations in this region fall into phenotypic classes which correspond to their map locations, suggesting that this region may consist of several distinct functional segments. Similarities between the organization of IS10's inverted repeat and those of other transposable elements are discussed. (ii) Base pairs 23-42 include a consensus binding sequence for one of the IS10 transposition host factors, IHF. The phenotypes of mutations in this region suggest that IHF is the major host factor for outside-end transposition activity in vivo and that base pairs throughout this region are important for the IHF interaction. (iii) Mutations in bp 43-61 do not affect outside-end transposition activity but do affect, in expected ways, previously identified determinants involved in expression and regulation of transposase. (iv) Some mutations in bp 23-42 also affect transposase expression; the possibility that IHF negatively regulates transcription initiation is discussed.
我们展示了对插入序列IS10外侧末端大量突变的遗传分析。(i) 末端反向重复序列可能是转座酶结合的主要位点。该区域的突变分为不同的表型类别,与它们在图谱上的位置相对应,这表明该区域可能由几个不同的功能区段组成。文中讨论了IS10反向重复序列的组织与其他转座元件的组织之间的相似性。(ii) 第23 - 42个碱基对包含IS10转座宿主因子之一IHF的共有结合序列。该区域突变的表型表明,IHF是体内外侧末端转座活性的主要宿主因子,并且该区域的所有碱基对对于IHF相互作用都很重要。(iii) 第43 - 61个碱基对中的突变不影响外侧末端转座活性,但确实以预期的方式影响先前确定的参与转座酶表达和调控的决定因素。(iv) 第23 - 42个碱基对中的一些突变也影响转座酶表达;文中讨论了IHF负调控转录起始的可能性。