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- W2494039341 abstract "The coordinated assembly of the DNA polymerase (gp43), the as( sliding clamp (gp45), and the clamp loader (gp44/62) to form the co bacteriophage T4 DNA polymerase holoenzyme is a multistep thi process. A partially opened toroid-shaped gp45 is loaded around m( DNA by gp44/62 in an ATP-dependent manner. Gp43 binds to this complex to generate the holoenzyme in which gp45 acts to ini topologically link gp43 to DNA, effectively increasing the procesgp sivity of DNA replication. Stopped-flow fluorescence resonance H( energy transfer was used to investigate the opening and closing of lo( the gp45 ring during holoenzyme assembly. By using two siteinl specific mutants of gp45 along with a previously characterized gp gp45 mutant, we tracked changes in distances across the gp45 74 subunit interface through seven conformational changes associOr ated with holoenzyme assembly. Initially, gp45 is partially open ho within the plane of the ring at one of the three subunit interfaces, in On addition of gp44/62 and ATP, this interface of gp45 opens ha further in-plane through the hydrolysis of ATP. Addition of DNA sai and hydrolysis of ATP close gp45 in an out-of-plane conformation. sh The final holoenzyme is formed by the addition of gp43, which re, causes gp45 to close further in plane, leaving the subunit interface lul open slightly. This open interface of gp45 in the final holoenzyme state is proposed to interact with the C-terminal tail of gp43, be providing a point of contact between gp45 and gp43. This study in further defines the dynamic process of bacteriophage T4 polymerarn ase holoenzyme assembly. ph gp" @default.
- W2494039341 created "2016-08-23" @default.
- W2494039341 creator A5018417584 @default.
- W2494039341 creator A5072073795 @default.
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- W2494039341 date "2016-01-01" @default.
- W2494039341 modified "2023-09-27" @default.
- W2494039341 title "Creating a dynamic picture o during T4 DNA polymerase h assembly by using fluorescen resonance energy transfer" @default.
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- W2494039341 hasPublicationYear "2016" @default.
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