Honigberg S M, Gonda D K, Flory J, Radding C M
J Biol Chem. 1985 Sep 25;260(21):11845-51.
Under conditions that diminish secondary structure in single-stranded DNA, stable presynaptic filaments can be formed by recA protein in the presence of the nonhydrolyzable analog ATP gamma S, without the need for Escherichia coli single strand binding protein. Such stable presynaptic filaments resemble those formed in the presence of ATP and pair efficiently with homologous duplex DNA. Since this kind of stable filament does not displace a strand from the duplex molecule, it provides a model substrate to study synapsis independent of the earlier and later stages of the recA reaction. Even though detectable strand displacement did not occur in the presence of ATP gamma S, both single strand and double strand breaks in duplex DNA stimulated homologous pairing. These and related observations support the view that the presynaptic nucleoprotein filament and naked duplex DNA intertwine to form a nascent joint in which the duplex DNA is partially unwound, i.e. in which the pitch of the involved duplex segment is reduced.
在能减少单链DNA二级结构的条件下,在不可水解的类似物ATPγS存在时,recA蛋白可形成稳定的突触前细丝,而无需大肠杆菌单链结合蛋白。这种稳定的突触前细丝类似于在ATP存在时形成的细丝,并能与同源双链DNA有效配对。由于这种稳定的细丝不会从双链分子中置换出一条链,因此它提供了一个模型底物,用于研究独立于recA反应早期和后期阶段的联会。尽管在ATPγS存在时未发生可检测到的链置换,但双链DNA中的单链和双链断裂均刺激同源配对。这些及相关观察结果支持这样一种观点,即突触前核蛋白细丝与裸露的双链DNA相互缠绕形成一个新生接头,其中双链DNA部分解旋,即其中所涉及双链片段的螺距减小。