Krawczyk B, Kur J
Department of Microbiology, Technical University of Gdańsk, Poland.
Acta Microbiol Pol. 1994;43(2):145-53.
The integration host factor (IHF) is a sequence-specific, histone-like, multifunctional DNA-binding and -bending protein of Escherichia coli. The characterization and functional analysis of this protein has been carried out mainly in bacteriophage lambda and other mobile genetic elements. Less is known concerning the role of IHF in E. coli, although it has been implicated in a number of processes in this organism including DNA replication, site-specific recombination, and gene expression. In this paper we report data concerning the binding of IHF protein to the recA gene region. IHF binds to at least four sites of the DNA fragment containing the recA gene, as shown by gel mobility shift assays. On the basis of the ihf consensus sequences published, we have been able to identify two sequences of putative ihf sites (ihf 1 and ihf2) into the 1390 bp long sequence containing the recA gene, but only the ihf2 site was able to bind IHF, as measured by gel mobility shift experiments. The nonfunctional ihf1 sequence was found to overlap the -35 region of the recA promoter and the functional ihf2 sequence was found within the recA gene structure at nt +780 to +807 (both with three mismatches according to the consensus sequence of Kur et al., 1989). This confirms our earlier results that the IHF-DNA interaction does not depend on any very rigid sequence, but also on the suitable sequences of the neighbouring regions, together with the proper DNA conformation.(ABSTRACT TRUNCATED AT 250 WORDS)
整合宿主因子(IHF)是大肠杆菌中一种序列特异性、组蛋白样、多功能的DNA结合和弯曲蛋白。对该蛋白的表征和功能分析主要在噬菌体λ和其他移动遗传元件中进行。尽管IHF已被认为参与了大肠杆菌中的许多过程,包括DNA复制、位点特异性重组和基因表达,但其在大肠杆菌中的作用仍知之甚少。在本文中,我们报告了有关IHF蛋白与recA基因区域结合的数据。凝胶迁移率变动分析表明,IHF与包含recA基因的DNA片段的至少四个位点结合。根据已发表的ihf共有序列,我们能够在包含recA基因的1390 bp长序列中鉴定出两个假定的ihf位点序列(ihf 1和ihf2),但通过凝胶迁移率变动实验测定,只有ihf2位点能够结合IHF。发现无功能的ihf1序列与recA启动子的-35区域重叠,而功能性的ihf2序列位于recA基因结构内的nt +780至+807处(根据Kur等人1989年的共有序列,两者均有三个错配)。这证实了我们早期的结果,即IHF与DNA的相互作用不仅取决于任何非常严格的序列,还取决于相邻区域的合适序列以及适当的DNA构象。(摘要截断于250字)