DNA-replication@生物化学精品讲义.pptxVIP

Happy Birthday,Double Helix ;DNA Replication;Molecular Structure of Nucleic Acids: A Structure for Deoxyribose Nucleic Acid (Nature, April 25, 1953. volume 171:737-738.) ;Testing Models for DNA replication;;Testing Models for DNA replication;Meselson and Stahl (continued);Meselson and Stahl Original Data;;;John Cairns;;DNA Replication is Semi-discontinuous;Evidence for the Semi-Discontinuous replication model was provided by the Okazakis (1968);Evidence for Semi-Discontinuous Replication (pulse-chase experiment) ;;;Enzymes and Proteins Involved in DNA Replication;DNA-dependent DNA polymerases;E. coli DNA polymerases;Arthur Kornberg (1957); How Amazing!!! a 3’ to 5’ exonuclease activity a 5’ to 3’ exonuclease activity a 5’ to 3’ DNA polymerizing activity ;;;Conceptual model for proofreading based on kinetic considerations;Proof reading activity of the 3’ to 5’ exonuclease. Proof reading activity is slow compared to polymerizing activity, but the stalling of DNAP I after insertion of an incorrect base allows the proofreading activity to catch up with the polymerizing activity and remove the incorrect base.;Notice how the newly-formed strand oscillates between the polymerase and 3-exonuclease sites, adding a base and then checking it;;;;;;;;;;The structure formed by two beta subunits of? the E. coli DNA polymerase III .? This structure can clamp a DNA molecule and slide with the core polymerase along the DNA molecule. ;;Comparison of E. Coli DNA pol I, II, and III;Eukaryotic DNA polymerase;Other Enzymes and Proteins Involved in DNA Replication;Action of Topoisomerase II ;Action of DNA Ligase;The “End-Replication Problem”;Act as protective “caps” on the ends of chromosomes. They are composed of short, tandem repeats. In humans: 5’-TTAGGG-3’ repeated at the ends of each chromosome for a total length of 15 kilobases. Telomeres are non-coding DNA Therefore, if telomeres gradually get eroded by DNA replication, there is less harm to the organism;Telo