Yurieva O, Skangalis M, Kuriyan J, O'Donnell M
Laboratories of Molecular Biophysics, The Rockefeller University, New York, New York 10021, USA.
J Biol Chem. 1997 Oct 24;272(43):27131-9. doi: 10.1074/jbc.272.43.27131.
This report identifies the dnaX homolog from Thermus thermophilis. Replicases from bacteria to humans contain subunits that are homologous to one another. These homologs are subunits of a clamp loading apparatus that loads sliding clamps onto DNA, which in turn act as mobile tethers for the replication machinery. In Escherichia coli, two of these subunits (gamma and tau) are encoded by one gene (dnaX) in nearly equal amounts by way of an efficient translational frameshift. The gamma and tau subunits form the central touchpoint that holds together two DNA polymerases with one clamp loading apparatus to form the E. coli chromosomal replicase, DNA polymerase III holoenzyme. The E. coli holoenzyme is an efficient replication machine that simultaneously replicates both strands of duplex DNA. The T. thermophilis dnaX homolog also contains a frameshift signature and produces both tau- and gamma-like proteins. Recombinant T. thermophilis tau- and gamma-like proteins, expressed in E. coli, have an oligomeric state similar to that of their E. coli counterparts and display ATPase activity that is stimulated by DNA. These results imply that T. thermophilis utilizes a DNA polymerase III holoenzyme replication machinery similar to that of E. coli.
本报告鉴定了嗜热栖热菌的dnaX同源物。从细菌到人类的复制酶都含有彼此同源的亚基。这些同源物是一种钳位装载装置的亚基,该装置将滑动钳装载到DNA上,而滑动钳又作为复制机器的移动系链。在大肠杆菌中,其中两个亚基(γ和τ)由一个基因(dnaX)通过高效的翻译移码以几乎相等的量编码。γ和τ亚基形成了一个中心接触点,将两个DNA聚合酶与一个钳位装载装置结合在一起,形成大肠杆菌染色体复制酶——DNA聚合酶III全酶。大肠杆菌全酶是一种高效的复制机器,能同时复制双链DNA的两条链。嗜热栖热菌的dnaX同源物也含有移码特征,并产生类似τ和γ的蛋白质。在大肠杆菌中表达的重组嗜热栖热菌τ和γ样蛋白质具有与其大肠杆菌对应物相似的寡聚状态,并表现出受DNA刺激的ATP酶活性。这些结果表明,嗜热栖热菌利用了一种与大肠杆菌类似的DNA聚合酶III全酶复制机制。