Wickner S, Hurwitz J
Proc Natl Acad Sci U S A. 1975 Sep;72(9):3342-6. doi: 10.1073/pnas.72.9.3342.
phiX174 DNA-dependent DNA synthesis is catalyzed in vitro by the combination of at least 11 purified protein fractions: dnaB, dnaC(D), and dnaG gene products, DNA polymerase III, DNA elongation factors I and II, DNA binding protein, and replication factors W, X, Y, and Z. The reaction requires ATP, 4 dNTPs, and Mg+2 and is specific for phiX174 (or phiXahb) DNA. Purified replication factor Y contains phiX174 (or phiXahb) DNA-dependent ATPase (or dATPase) activity. The ATPase activity is poorly stimulated by other single-stranded DNA, by double-stranded DNA, or by RNA. The products of the phiX174 DNA-dependent ATPase activity of factor Y are Pi and ADP (or dADP). The association of phiX174 DNA-dependent ATPase activity with factor Y was shown in the following ways: (a) the two activities copurified with a constant ratio; (b) they comigrated on native polyacrylamide gel electrophoresis; (c) both activities were heat-inactivated at the same rate; and (d) both showed identical patterns of N-ethylmaleimide sensitivity.
在体外，φX174 DNA依赖性DNA合成由至少11种纯化的蛋白质组分共同催化：dnaB、dnaC(D)和dnaG基因产物、DNA聚合酶III、DNA延伸因子I和II、DNA结合蛋白以及复制因子W、X、Y和Z。该反应需要ATP、4种脱氧核苷三磷酸和Mg²⁺，且对φX174（或φXahb）DNA具有特异性。纯化的复制因子Y具有φX174（或φXahb）DNA依赖性ATP酶（或dATP酶）活性。其他单链DNA、双链DNA或RNA对该ATP酶活性的刺激作用较弱。因子Y的φX174 DNA依赖性ATP酶活性的产物是无机磷酸和ADP（或dADP）。因子Y与φX174 DNA依赖性ATP酶活性的关联通过以下方式得以证明：(a) 两种活性以恒定比例共同纯化；(b) 它们在非变性聚丙烯酰胺凝胶电泳中迁移率相同；(c) 两种活性以相同速率被热灭活；以及(d) 两者对N-乙基马来酰亚胺的敏感性模式相同。
