Wiater L A, Grindley N D
Department of Molecular Biophysics and Biochemistry, Yale University, New Haven 06510.
EMBO J. 1988 Jun;7(6):1907-11. doi: 10.1002/j.1460-2075.1988.tb03024.x.
gamma delta, a prokaryotic transposon, encodes a transposase that is essential for its transposition. We show here, by DNase I protection experiments, that purified gamma delta transposase binds at the transposon's inverted repeats (IRs). Immediately adjacent to each transposase binding site (and within gamma delta DNA) we have identified a binding site for an additional protein factor, the Escherichia coli-encoded integration host factor (IHF). The binding of transposase and IHF to these adjacent sites is mutually cooperative. An IHF binding-site was also found in the original target DNA, just outside one of the ends of gamma delta. The affinity of IHF for this flanking site is reduced by transposase. These results demonstrate that gamma delta transposase binds at the IRs of gamma delta, and suggest that IHF may be involved in forming a transposase-DNA complex and/or influencing the target site selection during the transposition of gamma delta.
γδ是一种原核转座子,编码一种对其转座至关重要的转座酶。我们在此通过DNA酶I保护实验表明,纯化的γδ转座酶结合在转座子的反向重复序列(IRs)上。紧邻每个转座酶结合位点(且在γδ DNA内),我们鉴定出了另一种蛋白质因子——大肠杆菌编码的整合宿主因子(IHF)的结合位点。转座酶和IHF与这些相邻位点的结合是相互协同的。在γδ一端外侧的原始靶DNA中也发现了一个IHF结合位点。转座酶会降低IHF对这个侧翼位点的亲和力。这些结果表明γδ转座酶结合在γδ的IRs上,并提示IHF可能参与形成转座酶-DNA复合物和/或在γδ转座过程中影响靶位点选择。