DNAreplication08

DNAreplication08 - DNA REPLICATION Today's outline DNA...

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Julie Newdoll’s painting “Dawn of the Double Helix” Richard Dawkins wryly comments, “the individual is just DNA’s way of making more DNA!” DNA REPLICATION Today’s outline DNA synthesis: making a probe DNA synthesi,s DNA pols Structure, Mechanism Pol I, II and III Replication Other enzymes:
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DNA Synthesis: The Reaction Fig. 28-7, p. 905 DNA template with free 3’ OH Template = short ds region followed by ss region with free 3’OH To obtain: denature ds DNA, add ss DNA oligonucleotide primer Add: dNTPs Mg2+ DNA polymerase Nucleophilic attack by 3’O on α -phosphate of incoming dNTP Release of pyrophosphate- hydrolysis
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DNA Synthesis: The Reaction Fig. 28-7, p. 905 Incoming base is selected within DNA polymerase active site (WC base pair with template strand) Nucleophilic attack by 3’O on α -phosphate of incoming dNTP New phosphoester bond formed = chain growth in 5’ to 3’ direction (antiparallel to template strand) Release of PPi Subsequent hydrolysis of PPi by inorganic pyrophosphatase renders effectively irreversible
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A Universal Mechanism for All Polymerases Two metal ions on incoming nucleotide coordinated to phosphates Metal ions interact with D705 and D882 (T7 phage
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DNAreplication08 - DNA REPLICATION Today's outline DNA...

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