Wickner W, Brutlag D, Schekman R, Kornberg A
Proc Natl Acad Sci U S A. 1972 Apr;69(4):965-9. doi: 10.1073/pnas.69.4.965.
Soluble enzyme fractions from uninfected Escherichia coli convert M13 and varphiX174 viral single strands to their double-stranded replicative forms. Rifampicin, an inhibitor of RNA polymerase, blocks conversion of M13 single strands to the replicative forms in vivo and in vitro. However, rifampicin does not block synthesis of the replicative forms of varphiX174 either in vivo or in soluble extracts. The replicative form of M13 synthesized in vitro consists of a full-length, linear, complementary strand annealed to a viral strand. The conversion of single strands of M13 to the replicative form proceeds in two separate stages. The first stage requires enzymes, ribonucleoside triphosphates, and single-stranded DNA; the reaction is inhibited by rifampicin. The macromolecular product separated at this stage supports DNA synthesis with deoxyribonucleoside triphosphates and a fresh addition of enzymes; ribonucleoside triphosphates are not required in this second stage nor does rifampicin inhibit the reaction. We presume that in the first stage there is synthesis of a short RNA chain, which then primes the synthesis of a replicative form by a DNA polymerase.
来自未感染大肠杆菌的可溶性酶组分可将M13和φX174病毒单链转化为其双链复制形式。利福平是一种RNA聚合酶抑制剂，在体内和体外均可阻断M13单链向复制形式的转化。然而，利福平在体内或可溶性提取物中均不阻断φX174复制形式的合成。体外合成的M13复制形式由一条与病毒链退火的全长线性互补链组成。M13单链向复制形式的转化分两个独立阶段进行。第一阶段需要酶、核糖核苷三磷酸和单链DNA；该反应受利福平抑制。在该阶段分离出的大分子产物可支持用脱氧核糖核苷三磷酸和新添加的酶进行DNA合成；第二阶段不需要核糖核苷三磷酸，利福平也不抑制该反应。我们推测在第一阶段合成了一条短RNA链，然后由DNA聚合酶引发复制形式的合成。
