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Copyright © 2009 Pearson Education, Inc. PowerPoint ® Lecture Presentation for Concepts of Genetics Ninth Edition Klug, Cummings, Spencer, Palladino Chapter 11 DNA Replication and Recombination Lectures by David Kass with contributions from John C. Osterman. Copyright © 2009 Pearson Education, Inc.
DNA Is Reproduced by Semiconservative Replication The complementarity of DNA strands allows each strand to serve as a template for synthesis of the other. Section 11.1
Copyright © 2009 Pearson Education, Inc. 3 possible modes of DNA replication are possible: conservative semiconservative dispersive Section 11.1
Copyright © 2009 Pearson Education, Inc. The Meselson-Stahl experiment demonstrated that: DNA replication is semiconservative each new DNA molecule consists of one old strand and one newly synthesized strand Section 11.1
Copyright © 2009 Pearson Education, Inc. Figure 11.3
Copyright © 2009 Pearson Education, Inc. Figure 11.4
Copyright © 2009 Pearson Education, Inc. Meselson-Stahl Experiment
Copyright © 2009 Pearson Education, Inc. The Taylor- Woods-Hughes experiment demonstrated that DNA replication is semiconservative in eukaryotes. Section 11.1
Copyright © 2009 Pearson Education, Inc. Semiconservative Replication - Mechanism of DNA Replication (Basic) - Mechanism of DNA Replication (Advanced) - DNA Replication Process -
Copyright © 2009 Pearson Education, Inc. DNA replication begins at the origin of replication and is bidirectional rather than unidirectional. A replicon is the length of DNA that is replicated following one initiation event at a single origin. Section 11.1
Copyright © 2009 Pearson Education, Inc. DNA Synthesis in Bacteria Involves Five Polymerases, as well as Other Enzymes DNA polymerase I catalyzes DNA synthesis and requires a DNA template and all four dNTPs. Section 11.2
Copyright © 2009 Pearson Education, Inc. Chain elongation occurs in the 5' to 3' direction by addition of one nucleotide at a time to the 3' end. Polymerase Direction
Copyright © 2009 Pearson Education, Inc. DNA polymerases I, II, and III can elongate an existing DNA strand (called a primer) but cannot initiate DNA synthesis. All three possess 3' to 5' exonuclease activity. But only DNA polymerase I demonstrates 5' to 3' exonuclease activity. Section 11.2
Copyright © 2009 Pearson Education, Inc. DNA polymerase III is the enzyme responsible for the 5' to 3' polymerization essential in vivo. Its 3' to 5' exonuclease activity allows proofreading. Section 11.2
Copyright © 2009 Pearson Education, Inc. Polymerase I is believed to be responsible for: removing the primer the synthesis that fills gaps produced during synthesis Section 11.2
Copyright © 2009 Pearson Education, Inc. DNA polymerases I, II, IV, and V are involved in various aspects of repair of damaged DNA. Section 11.2
Copyright © 2009 Pearson Education, Inc. DNA polymerase III has 10 subunits whose functions are shown in Table Section 11.2
Copyright © 2009 Pearson Education, Inc. Polymerase III Holoenzyme (made of many protein subunits) in E. coli Shevelev, Igor and Hubschur, Ulrich “The 3’ to 5’ exonucleases.” Nature Reviews Molecular Cell Biology 3, pg Retrieved 11/5/13 from
Copyright © 2009 Pearson Education, Inc. 7 key issues that must be resolved during DNA replication: unwinding of the helix reducing increased coiling generated during unwinding synthesis of a primer for initiation discontinuous synthesis of the second strand removal of the RNA primers joining of the gap-filling DNA to the adjacent strand proofreading Section 11.3 Many Complex Tasks Must Be Performed during DNA Replication
Copyright © 2009 Pearson Education, Inc. DnaA binds to the origin of replication (oriC) and is responsible for the initial steps in unwinding the helix. Section 11.3 – Unwinding DNA Helix
Copyright © 2009 Pearson Education, Inc. To elongate a polynucleotide chain, DNA polymerase III requires a primer with a free 3'- OH group. Enzyme primase synthesizes an RNA primer that provides the free 3'-OH required by DNA polymerase III Section RNA Primer
Copyright © 2009 Pearson Education, Inc. As replication fork moves, only 1 strand can serve as template for continuous DNA synthesis—the leading strand. Opposite lagging strand undergoes discontinuous DNA synthesis. Section 11.3
Copyright © 2009 Pearson Education, Inc. Both DNA strands are synthesized concurrently by looping the lagging strand to invert the physical but not biological direction of synthesis. Section 11.3
Copyright © 2009 Pearson Education, Inc. Proofreading and error correction are an integral part of DNA replication. All of the DNA polymerases have 3' to 5' exonuclease activity that allows proofreading. Section 11.3
Copyright © 2009 Pearson Education, Inc. DNA synthesis at a single replication fork: Section 11.4
Copyright © 2009 Pearson Education, Inc. Eukaryotic DNA Synthesis Is Similar to Synthesis in Prokaryotes, but More Complex In eukaryotic cells: there is more DNA than prokaryotic cells the chromosomes are linear the DNA is complexed with proteins Section 11.6
Copyright © 2009 Pearson Education, Inc. Eukaryotic chromosomes contain multiple origins of replication to allow the genome to be replicated in a few hours. Section 11.6
Copyright © 2009 Pearson Education, Inc. 3 DNA polymerases are involved in replication of nuclear DNA. 1 involves mitochondrial DNA replication. Others are involved in repair processes. Section 11.6
Copyright © 2009 Pearson Education, Inc. Pol and and Ɛ major forms of the enzyme involved in initiation and elongation. Pol possesses low processivity. functions in synthesis of RNA primers during initiation on the leading and lagging strands. Polymerase switching occurs Pol is replaced by Pol and Ɛ which has high processivity, for elongation. Section 11.6
Copyright © 2009 Pearson Education, Inc. Telomeres Provide Structural Integrity at Chromosome Ends but Are Problematic to Replicate Telomeres at the ends of linear chromosomes consist of long stretches of short repeating sequences and preserve the integrity and stability of chromosomes. Section 11.7
Copyright © 2009 Pearson Education, Inc. T-Loop in Telomeres
Copyright © 2009 Pearson Education, Inc. T-Loop - Telomere
Copyright © 2009 Pearson Education, Inc. Lagging strand synthesis at end of chromosome is a problem b/c once the RNA primer is removed, there is no free 3'-hydroxyl group from which to elongate. Section 11.7
Copyright © 2009 Pearson Education, Inc. Telomerase directs synthesis of the telomere repeat sequence to fill gap. This enzyme is a ribonucleoprotein w/an RNA that serves as the template for the synthesis of its DNA complement. Section 11.7
Copyright © 2009 Pearson Education, Inc. DNA Recombination, Like DNA Replication, Is Directed by Specific Enzymes Genetic recombination involves: endonuclease nicking strand displacement ligation branch migration duplex separation to generate the characteristic Holliday structure (chi form) Section 11.8
Copyright © 2009 Pearson Education, Inc. Figure 11.18
Copyright © 2009 Pearson Education, Inc. Gene conversion is characterized by nonreciprocal genetic exchange between two closely linked genes. Section 11.9 Gene Conversion Is a Consequence of DNA Recombination
Copyright © 2009 Pearson Education, Inc. The End
Copyright © 2009 Pearson Education, Inc. PowerPoint ® Lecture Presentation for Concepts of Genetics Ninth Edition Klug, Cummings, Spencer, Palladino Chapter.
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