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DNA Replication
DNA is semiconservative o Semiconservative Semiconservative replication: A replication: A mechanism of DNA replication in which each of the two strands of parent DNA is incorporated into a new double stranded DNA molecule. Involves separating the two parent strand and building a new o complementary replacement strand for each
DNA Replication: The process process Step 1: Strand Separation DNA must first unwound each other (replication origin is the starting point) o Replication Origin: A Origin: A specific sequence of DNA that acts as a starting point for replication Helicase binds to the starting point and starts to unwind the two DNA strands by breaking the hydrogen bonds between the complementary base pairs. Helicase: A Helicase: A replication enzyme that separates and unwinds the DNA o strands As strands start to separate, they for a y shape known as the replication fork. o Replication Fork: The point of separation of the two parent DNA DNA strands during replication An enzyme called Topoisomerases prevents DNA strands from being twisted and tangled. It relieves tension by cutting one or two of the strand near the o replication fork Topoisomerase: An Topoisomerase: An enzyme that relieve tension caused by the o unwinding of parent DNA; they cleave one or two of the DNA strand, st rand, allow the strands to untwist and then rejoin the strands DNA has the tendency to separate and a nd bind together again so to prevent that SSBs prevents the H bonds from reforming and annealing. o SSBs (single-strand binding protein): A protein): A replication enzyme that prevents parent DNA strand from annealing to each other once they have been separated by helicase
Step 2: Building Complementary Strands New nucleotides are joined by a group of enzymes called DNA polymerase. Both prokaryotes and eukaryotes have DNA polymerase Prokaryotic cell add DNA from 3’ to 5’ end while eukaryotic cell add DNA from 5’ to 3’ end Builds new strand by using nucleoside triphosphates o a building block and energy source for replicating DNA RNA primase begins the replication stage and set small complementary RNA segment at the beginning of the replication fork
RNA Primase: A replication enzyme that produces RNA primers About 10-60 RNA are produced also known as RNA primer RNA Primer: A replication molecule that acts as a starting point for o replication Once RNA primer is set, DNA polymerase III starts adding DNA nucleotides to the RNA primer o DNA polymerase III: A prokaryotic replication enzyme that builds new DNA strands from nucleotides Can only build from 5’ to 3’ end o Strands start building in opposite direction The DNA polymerase III that is building towards the replication for is able to continuously build (no RNA Primer needs to keep adding to RNA primase) o Known as the Leading strand o Leading Strand: The DNA strand that is copied in the direction towards the replication fork On the other strands, the DNA polymerase III is replicating away from the replicating fork. o Lagging Strand: The DNA strand that is copied in the direction away from the replication fork RNA Primase will attach itself to part of the lagging strand and will be replicating in pieces called Okazaki fragments o Okazaki fragments: The piece of new DNA on the lagging strand Lagging strand built in pieces and is not a continual process since it is made in pieces Each fragment extend in the 5’ to 3’ direction, it eventually runs into the RNA primer attached to the Okazaki fragment ahead DNA polymerase I removes the RNA primer/nucleotides one at a time and replaces it with DNA nucleotides o DNA Polymerase I: A prokaryotic replication enzyme that fills in gaps in the lagging strand between Okazaki fragments; also proofreads the final strand DNA ligase connects the new DNA nucleotides and Okazaki fragments together o DNA Ligase: An enzyme that catalyzes the formation of a phosphodiester bond between two DNA strands, as well as between Okazaki fragment. o
Step 3: Dealing with Errors during DNA Replication A DNA polymerase proof reads the strand and corrects the errors (usually base pair match) DNA Polymerase III cannot move on if there is a error in the strand Composed of proteins and enzymes including DNA polymerase I and polymerase II DNA polymerase II is a slow enzyme that works to repair damages to DNA
DNA polymerase II : A prokaryotic replication enzyme that repairs damage to DNA, including damage that occurs between replication event Mismatch base pairs will not be able to bond correctly and will distort the shape of the DNA o
