Groenen M A, Vollering M, Krijgsman P, van Drunen K, van de Putte P
Department of Molecular Genetics, University of Leiden, The Netherlands.
Nucleic Acids Res. 1987 Nov 11;15(21):8831-44. doi: 10.1093/nar/15.21.8831.
Transposition of the E. coli bacteriophage Mu requires the phage encoded A and B proteins, the host protein HU and the host replication proteins. The ends of the genome of the phage, on which some of these proteins act, both contain three transposase (A) binding sites. The organization of these binding sites on each end, however, is different. Here we show, using DNase footprinting experiments with purified A protein, that mutant A binding sites, which affect transposition, have decreased affinity for the transposase. Furthermore the transposase binds non-cooperatively to all A binding sites both in the left and right end of Mu. Electron microscopic studies show that the A protein forms specific nucleoprotein structures upon binding to the ends of Mu. The A and B proteins interact with the ends of Mu to generate larger structures than with the A protein alone.
大肠杆菌噬菌体Mu的转座需要噬菌体编码的A和B蛋白、宿主蛋白HU以及宿主复制蛋白。这些蛋白作用于噬菌体基因组的末端,两端均含有三个转座酶(A)结合位点。然而,两端这些结合位点的组织方式不同。在这里,我们通过使用纯化的A蛋白进行DNA酶足迹实验表明,影响转座的突变型A结合位点对转座酶的亲和力降低。此外,转座酶与Mu左端和右端的所有A结合位点均非协同结合。电子显微镜研究表明,A蛋白在与Mu末端结合时形成特定的核蛋白结构。A和B蛋白与Mu末端相互作用,产生比单独与A蛋白作用时更大的结构。