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Simultaneous Formation Of Functional Leading And Lagging Strand Holoenzyme Complexes On A Small, Defined Dna Substrate, Anthony J. Berdis, Stephen J. Benkovic
Simultaneous Formation Of Functional Leading And Lagging Strand Holoenzyme Complexes On A Small, Defined Dna Substrate, Anthony J. Berdis, Stephen J. Benkovic
Chemistry Faculty Publications
The biochemical characterization of leading and lagging strand DNA synthesis by bacteriophage T4 replication proteins has been addressed utilizing a small, defined primer/template. The ATP hydrolysis activity of 44/62, the clamp loading complex responsible for holoenzyme assembly, was monitored during assembly of both the leading and lagging strand holoenzyme complex. The ATPase activity of 44/62 diminishes once a functional holoenzyme is assembled on both the leading and lagging strand. The assembly of the lagging strand holoenzyme is facilitated by several factors including biotinylated streptavidin blocks at the end of the fork strands, preassembly of the leading strand holoenzyme, and by …