DNA REPLICATION AND REPAIR before cell division occurs the DNA molecule unwinds and is replicated DNA Replicates Semiconservatively each daughter cell receives one strand from the parent molecule and one strand is newly synthesized see Fig. 1 on pg. 217 Matthew Meselson and Franklin Stahl devised an experiment to prove this in 1958 The Process of DNA Replication Separating the DNA Strands replication begins when a protein binds to a specific site on the DNA molecule called the replication origin the linear DNA of eukaryotes have more than one replication origin, while the DNA of prokaryotes have only one an enzyme (DNA helicase) is responsible for breaking the hydrogen bonds between the nitrogenous base pairs causing the helix to unwind single stranded binding proteins binds to the exposed bases and prevent hydrogen bonding from occurring during bacterial replication an enzyme called DNA gyrase cuts the strands to relieve the tension from unwinding then reseals them later DNA replication will begin at the origins and move out in both directions replication forks will exist where the two strands are still joined one strand moves towards the fork while the other moves away eukaryotes contain many forks since they have multiple sites or origin a replication bubble forms between two closely situated forks see Fig. 4 on pg. 220 Building the Complementary Strand DNA polymerase III is the enzyme that builds the complementary strand an RNA primer (a sequence of 10 – 60 RNA bases) sticks to the 3’ end with the help of an enzyme called primase it recognizes the 3’ end and adds a deoxyribonucleoside triphosphate to the end DNA polymerase III is now free to add free deoxyribonucleotide triphosphates the enzyme uses the energy from breaking the bond between the first and second phosphate group so carry out the condensation reaction that is responsible for adding the complementary nucleotide to the strand the two remaining phosphate are recycled to make more nucleoside triphosphates the 3’ to 5’ strand is the template used as a guide to create the leading strand (it moves towards the fork), and builds continuously the 5’ to 3’ strand is the template used as a guide to create the lagging strand (it moves away from the fork), and is built discontinuously DNA is built in short segments (Okazaki fragments) by Polymerase III DNA Polymerase I removes the RNA primers in both the leading and lagging strands and replace them with the appropriate deoxyribonucleotides. DNA ligase joins the Okazaki fragments on the lagging strands by creating a phosphodiester bond See Fig. 7 on pg. 221 resulting in two double-stranded DNA molecules that automatically form a helix. DNA Repair DNA polymerase III and I act as quality control checkers by proofreading the new strands if there is a mistake the enzyme can function as an exonuclease; an enzyme that is free to cut out nucleotides that are incorrectly paired, and add new ones. Seatwork Pg. 223 # 1, 2, 4 – 7, 8, 10