Hwang E S, Scocca J J
Department of Biochemistry, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205.
J Bacteriol. 1990 Sep;172(9):4852-60. doi: 10.1128/jb.172.9.4852-4860.1990.
The specific DNA-binding protein integration host factor (IHF) of Escherichia coli stimulates the site-specific recombination reaction between the attP site of bacteriophage HP1 and the attB site of its host, Haemophilus influenzae, in vitro and also appears to regulate the expression of HP1 integrase. IHF interacts specifically with DNA segments containing the att sites and the integrase regulatory region, as judged by IHF-dependent retardation of relevant DNA fragments during gel electrophoresis. The locations of the protein-binding sites were identified by DNase I protection experiments. Three sites in the HP1 attP region bound IHF, two binding sites were present in the vicinity of the attB region, and one region containing three partially overlapping sites was present in the HP1 integrase regulatory segment. The binding sites defined in these experiments all contained sequences which matched the consensus IHF binding sequences first identified in the lambda attP region. An activity which stimulated the HP1 site-specific integration reaction was found in extracts of H. influenzae, suggesting that an IHF-like protein is present in this organism.
大肠杆菌的特异性DNA结合蛋白整合宿主因子(IHF)在体外可刺激噬菌体HP1的attP位点与其宿主流感嗜血杆菌的attB位点之间的位点特异性重组反应,并且似乎还调控HP1整合酶的表达。通过凝胶电泳过程中相关DNA片段的IHF依赖性阻滞判断,IHF与含有att位点和整合酶调控区域的DNA片段特异性相互作用。通过DNase I保护实验确定了蛋白质结合位点的位置。HP1 attP区域有三个位点结合IHF,attB区域附近有两个结合位点,并且在HP1整合酶调控片段中有一个包含三个部分重叠位点的区域。在这些实验中确定的结合位点均包含与最初在λattP区域中鉴定出的共有IHF结合序列相匹配的序列。在流感嗜血杆菌的提取物中发现了一种刺激HP1位点特异性整合反应的活性,这表明该生物体中存在一种类似IHF的蛋白质。