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ISU BBMB 405 - DNA Replication
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BBMB 405 1st Edition Lecture 23 Outline of Last Lecture XVIII. Chapter 26: The Biosynthesis of Membrane Lipids and SteroidsD. Important derivatives of cholesterol include bile salts and steroid hormonesOutline of Current Lecture XIV. Chapter 28: DNA Replication, Repair and RecombinationA. Introduction: Central Dogma and ReviewB. DNA replication proceeds by polymerization of deoxyribonucleoside triphosphates along templateCurrent LectureXIV. Chapter 28: DNA Replication, Repair and RecombinationA. Introduction: Central Dogma and Review1. DNA transcribed to RNA is translated to Protein2. Types of polymerases: a. DNA dependent DNA polymerase: replicationb. DNA dependent RNA polymerase: transcriptionc. RNA dependent DNA polymerase: reverse transcriptiond. RNA dependent RNA polymerase: RNA replication3. Requirements of polymerases? Template, dNTPs or NTPs, metal ions, DNA also requires a primerThese notes represent a detailed interpretation of the professor’s lecture. GradeBuddy is best used as a supplement to your own notes, not as a substitute.4. DNA structure review: right handed helix, 10 to 10.5 pairs per turn, minor grove (lead to non specific reacitons) and major grove, directionality 5’ to 3’5. DNA replication is semi conservative6. What are requirements for DNA replication? Template, speed/processivity, fidelity, repair7. DNA must be accurately replicated and maintained1. Errors can occur during replication via incorrect incorporation2. DNA damage can occur at any time3.8. DNA polymerase review: need to have triphosphate because if primary phosphate is hydrolyzed then polymerase stallsB. DNA replication proceeds by polymerization of deoxyribonucleoside triphosphates along template1.2. Fingers are nucleotide binding, ssDNA template interaction3. Palm has catalytic residues and is where DNA sits4. Thumb is dsDNA product interaction5. Two metal ion mechanism of DNA polymerase; metal is activated nucleophile for attack on phosphate6. Considerations for fidelity of DNA polymerasea. Nucleotide misincorporation: errors occur at rate of 10^-5 bp for DNA polymeraseIIIb. Mismatch Extension: errors are missed by DNA pol III proofreading at rate of 10^-2 bpc. Overall error rate of DNA pol III is 10^-7 bp7. What determines if aquired correct nucleotide for binding? Shape of nucleotide8. Why shapte complementarity so important? Residues (Arg and Gln) of enzyme form hydrogen bonds in minor groove side of base pair in active site. The hydrogen bond acceptors act as ruler that measure if properly spaced base pair has formed in active site. DNA polymerases close down around incoming dNTP, the binding triggers conformational change where finger domain rotates to form tight pocket into which only properly shaped base pair fits9. Model of DNA binding to two active sitesa. Primer strand can transfer between polymerase and 3’-5’-exonuclease active sitesb. Destabilized dsDNA favors editing complex10. Note: rest of notes from this lecture overlap with next lecture (if they are not in this lecture’s notes they are in next lecture’s


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ISU BBMB 405 - DNA Replication

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