Lecture 5: DNA Replication - Elongation part 1
1. Helicase: a. What does it do in DNA replication? b. Does it require ATP - if so why, and if not why not? c. Looking at one replication fork, does helicase travel on the leading-strand-template or the lagging-strand-template? Why is it important that it be on this strand (think about another enzyme helicase is associated with)?
2. Single-Stranded DNA Binding Protein (SSBP): a. What does it do in DNA replication? b. What part(s) of the DNA strand does this protein recognize for binding?
3. Topoisomerase II: a. What is another name for this enzyme? b. What does it do in DNA replication, and through what activity does it accomplish this? c. Does it require ATP - if so why, and if not why not?
4. Primase: a. What does it do in DNA replication? b. Does it require ATP - if so why, and if not why not? c. Looking at one replication fork, does Primase travel on the leading-strand-template or the lagging-strand-template? Why is it important that it be on this strand? d. What other enzyme is Primase attached to, and when does it attach to it? e. Primase creates new polynucleotides in the ____' to ____' direction. f. Which nucleotide would Primase recognize as a substrate: dCTP or CTP? Why? g. Is Primase a type of DNA Polymerase or a type of RNA Polymerase?
5. DNA Polymerase III: a. What does it do in DNA replication? b. Does it require ATP - if so why, and if not why not? c. DNA Pol III reads the parental template strand in the ____' to ____' direction, and creates the new daughter strand in the ____' to ____' direction. d. Which nucleotide would DNA Pol III recognize as a substrate: dCTP or CTP? Why? e. Why are primers important to DNA Pol III? f. DNA Pol III creates a covalent bond between the ____'-____________ group of the new nucleotide and the ____'-____________ group of the already existing DNA strand (or of the primer, if just starting to make DNA). g. DNA Pol III has two enzymatic activities: ____' to ____' ____________ase activity and ____' to ____' ____________ase activity. Which of these activities is part of the enzyme's proofreading function? h. What is meant by proofreading?