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3. Replication - DNAreplica,oncomputeranima,on Replica,on...

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DNA replica,on‐computer anima,on
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Replica,on: I: Ge7ng started II: Elonga,on II: Telomerase and the end game
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What is an origin? In bacteria it is a specific DNA sequence, oriC and is unique in the chromosome. What specific proper,es does it need?
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Separa,ng strands of duplex DNA requires energy Energy What features of an origin facilitate stand separa,on in vivo, ie, how do cells reduce the energy barrier of strand separa,on?
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It uses the energy of ATP binding to wrap DNA in posi,ve supercoils. This induces local nega,ve supercoiling in the A:T‐rich DNA unwinding element, facilita,ng “mel,ng” of the origin. DNA unwinding element DnaA is a AAA+ ATPase that binds DNA. DnaA monomer DnaA filament
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Figure 06-21 DnaC then recruits DnaB, the replica,ve helicase
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Figure 06-11 The replica,ve helicase from Papilloma virus, another AAA+ ATPase ATP binding site The central channel can accommodate only a single strand of DNA
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Biochemical assay for DNA helicase activity Karow J K et al. J. Biol. Chem. 1997;272:30611-30614 ©1997 by American Society for Biochemistry and Molecular Biology
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Finding a needle in a hay stack How do you figure out what defines a eukaryo,c origin of replica,on?
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