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Lecture11CMFall09FINAL - One week volunteer trip in...

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One week volunteer trip in Honduras Second week of summer (May 24) GWB goes to source to find and implement clean water and sanitation solutions Talk with locals about clean water and proper hygiene Spanish background is beneficial First meeting October 1at 7:00 pm in 213 Greg Hall Contact: Chris [email protected] Scott [email protected] http://www.globalbrigades.org/project/water/
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MCB 250 Lecture 11 DNA Replication in Bacteria (Continued)
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DNA replication Pol I and RnaseH remove RNA primer completing DNA Synthesis DNA Ligase Seals Nicks Okazaki Fragments Replisome (2xPol III) Synthesizes both Leading and Lagging Strands at Replication Fork 5’ 3’ 5’ 3’ P OH Nick 5’ 3’ 5’ 3’
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Travels 5’ 3’ On the lagging strand Uses energy from ATP to unwind the helix Text Fig 8-13 A Helicase is Required to Unwind the DNA Duplex
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Text Fig. 8-15 Binding of SSB is highly cooperative. Binding of one SSB makes it likely that another will bind next to it. A single strand binding protein (SSB) binds to single stranded regions formed by helicase to prevent reformation of the double helix. Single Strand Binding Protein
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Proteins Required for Replication Helicase (DnaB) Melts parental DNA, interacts with PolIII and Primase Primase (DnaG) Synthesizes RNA primers DNA Pol III Synthesizes DNA by adding bases to a primer SSB (Single Strand Binding Protein) Binds single stranded DNA template cooperatively, prevents reannealing and hairpins RNAse H Removes RNA primers Pol I Removes RNA primers and fills gap with DNA DNA ligase Seals nicks between Okazaki fragments
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The Replisome Leading Strand DNA Polymerase Lagging Strand DNA Polymerase Sliding Clamp γ -Complex τ -Protein DNA Helicase SSB Bound To DNA 5’ 3’ 3’ 5’ 5’ 3’ 5’ 3’ 5’ 3’OH Primase Text Fig. 8-22
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The Cast of Characters The star of the show DNA polymerase III (PolIII)
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E. coli DNA Polymerase III (PolIII) Core polymerase - a heterotrimer α – DNA Polymerase ε – 3’ to 5’ Exonuclease θ – Stimulates Exonuclease “Holoenzyme” is composed of 10 different subunits 2 Core polymerases 2(
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