Objectives Unit Nine:  DNA Replication, Synthesis and Recombination

1.  Define the following terms and utilize these terms in an appropriate context:
bidirectional replication             conservative replication                  continuous DNA synthesis
dispersive replication                lagging strand                                   leading strand
nucleosome                               Okazaki fragment                              ori
origin of replication                    primer                                                replication fork
replicon                                       semiconservative replication

2.  Compare conservative, semiconservative and dispersive modes of DNA replication.

3.  Summarize the properties and functions of polymerase I, II and III.

4.  Discuss the roles of the polymerase molecules in maintaining the fidelity of the replication process.

5.  Define and indicate the significance of  A. Okazaki fragments,  B. DNA ligase,  C. primer RNA during DNA synthesis.

6.  Outline the current model for DNA synthesis.

7.  Compare and contrast DNA synthesis in prokaryotes and eukaryotes.

8.  Show by diagrams how a phosphate group joins the 3' carbon of one deoxyribose to the 5' carbon of the next deoxyribose.

9.  Describe the various characteristics of the Watson-Crick double-helix model for DNA.

10.  Explain the statement that DNA consists of antiparallel and complementary strands, which read 3'-5' on one strand, but '5-3' on the other.

11.  Describe the bonding between the nucleotide bases on one DNA strand and the nucleotide bases of the antiparallel strand.

Resources:  Text Chapter 10, Cartoon Guide pgs. 104-107; 125-128
 

DNA/RNA Synthesis Replication

Replication Origins
    -is there a single origin or many?
    -origin random or in a specific region?

        Replicon- length of DNA replicated following initiation of synthesis at a single origin

        Replication fork- point of replication, where strand is unwinding

    Bacteria/viruses: single chromosomes specific region of initiation (ori) entire chromosome one replicon
    Eukaryotes: replication bidirectional
        2 replication forks, multiple origins along chromosome (eventually merge)

        multiple replicons (because of length allows for quicker replication)

DNA Synthesis: A model
    Semiconservative; bidirectional

    1. mechanism by which helix is denatured and stabilized so that replication can occur in both directions
            -helicase
            -single strand-DNA binding protein (binds to newly seperated DNA  strands to prevent reassociation)

    2. As unwinding proceeds tension created, must be reduced
            -DNA gyrase (DNA topoisomerase), causes break in both strands which is resealed after tension
            lessened

    3. Primer created so polymerization can commence under direction of DNA polymerase (requires a free 3'
        end)
            -RNA primase
            -DNA polymerase adds 5" nucleotides

    4. Synthesis of complementary strands
                Strands are antiparallel
                    5' ---> 3'
                    3' <--- 5'
            -continuous replication in the direction of replication fork possible only on one of two strands

            -other strand discontinuous in opposite direction

                    Leading strand- strand of continuous replication
                    Lagging strand- strand of discontinuous replication; Many points of initiation

                   Okazaki fragments- short fragments of DNA 1000-2000 nucleotides long that are enzymatically
                    joined to create longer DNA strand
                        -occur on lagging strand

    5. RNA primers must be removed
            DNA ligase- enzyme that can unite smaller fragments of DNA
            -joins gaps created by removal of primer
            -joins Okazaki fragments.

    6. Replication occurs along length of replicon

Fidelity of Synthesis

    Complicated processes need safeguards
        -RNA primer
        -discontinuous replication
        -semiconservative replication
        -complementary base pairing

     In addition:
        -Small alphabet of 4 letters
        -very few rules ---> A-T; C-G

        DNA polymerases- proofreading activity
            -will discontinue synthesis if previous base pair is mismatched

    Genes have been identified for the production of products involved in:
        -origin of synthesis
        -helix unwinding and stablization
        -initiation and priming
        -relaxation of supercoiling
        -repair and ligation
 

    Recombination at the molecular level
        -crossing over in meiosis can be between two seperate DNA molecules (usually homologous) or
        intramolecular

        -genetic recombination

    Models depend on:
        1. 2 homologous double-stranded DNA molecules
        2. Complementarity between DNA strands
        3. series of enzymatic processes to accomplish recombination, pg 188

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