1 DNA and Replication

2 DNA Structure

3 DNA Stands for Deoxyribonucleic acid nucleotidesMade up of subunits called nucleotides Nucleotide made of:Nucleotide made of: Phosphate group 1.Phosphate group 5-carbon sugar 2.5-carbon sugar Nitrogenous base 3.Nitrogenous base

4 DNA Nucleotide O=P-O OPhosphate Group Group N Nitrogenous base (A, G, C, or T) (A, G, C, or T) CH2 O C1C1 C4C4 C3C3 C2C2 5 Sugar Sugar(deoxyribose) O

5 Pentose Sugar Carbons are numbered clockwise 1’ to 5’ CH2 O C1C1 C4C4 C3C3 C2C2 5 Sugar Sugar(deoxyribose)

6 DNA P P P O O O P P P O O O G C TA

7 Antiparallel Strands One strand of DNA goes from 5’ to 3’ (sugars) The other strand is opposite in direction going 3’ to 5’ (sugars)

8 Nitrogenous Bases Double ring PURINESDouble ring PURINES Adenine (A) Guanine (G) Single ring PYRIMIDINESSingle ring PYRIMIDINES Thymine (T) Cytosine (C) T or C A or G

9 Base-Pairings Purines only pair with Pyrimidines Three hydrogen bonds required to bond Guanine & Cytosine CG 3 H-bonds

10 T A Two hydrogen bonds are required to bond Adenine & Thymine

11 Question: Adenine CytosineIf there is 30% Adenine, how much Cytosine is present?

12 Answer: CytosineThere would be 20% Cytosine Adenine (30%) = Thymine (30%)Adenine (30%) = Thymine (30%) Guanine (20%) = Cytosine (20%)Guanine (20%) = Cytosine (20%) Therefore, 60% A-T and 40% C-GTherefore, 60% A-T and 40% C-G

13 DNA Replication

14 Replication Facts DNA has to be copied before a cell dividesDNA has to be copied before a cell divides DNA is copied during the S or synthesis phase of interphaseDNA is copied during the S or synthesis phase of interphase New cells will need identical DNA strandsNew cells will need identical DNA strands

15 Synthesis Phase (S phase) S phase during interphase of the cell cycle Nucleus of eukaryotes Mitosis -prophase -metaphase -anaphase -telophase G1G1 G2G2 S phase interphase DNA replication takes place in the S phase.

16 DNA Replication Begins at Origins of ReplicationBegins at Origins of Replication Two strands open forming Replication Forks (Y-shaped region)Two strands open forming Replication Forks (Y-shaped region) New strands grow at the forksNew strands grow at the forks ReplicationFork Parental DNA Molecule 3’ 5’ 3’ 5’

17 DNA Replication As the 2 DNA strands open at the origin, Replication Bubbles formAs the 2 DNA strands open at the origin, Replication Bubbles form Prokaryotes (bacteria) have a single bubble Eukaryotic chromosomes have MANY bubbles Bubbles

18 DNA Replication Enzyme Helicase unwinds and separates the 2 DNA strands by breaking the weak hydrogen bondsEnzyme Helicase unwinds and separates the 2 DNA strands by breaking the weak hydrogen bonds Single-Strand Binding ProteinsSingle-Strand Binding Proteins attach and keep the 2 DNA strands separated and untwisted

19 DNA Replication Before RNA primersBefore new DNA strands can form, there must be RNA primers present to start the addition of new nucleotides PrimasePrimase is the enzyme that synthesizes the RNA Primer DNA polymerase can then add the new nucleotides

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21 DNA Replication DNA polymerase can only add nucleotides to the 3’ end of the DNADNA polymerase can only add nucleotides to the 3’ end of the DNA This causes the NEW strand to be built in a 5’ to 3’ directionThis causes the NEW strand to be built in a 5’ to 3’ direction RNAPrimer DNA Polymerase Nucleotide 5’ 3’ Direction of Replication

22 Remember HOW the Carbons Are Numbered! O O=P-O OPhosphate Group Group N Nitrogenous base (A, G, C, or T) (A, G, C, or T) CH2 O C1C1 C4C4 C3C3 C2C2 5 Sugar Sugar(deoxyribose)

23 Remember the Strands are Antiparallel P P P O O O P P P O O O G C TA

24 Synthesis of the New DNA Strands The Leading Strand single strandThe Leading Strand is synthesized as a single strand from the point of origin toward the opening replication fork RNAPrimer DNA Polymerase Nucleotides 3’5’

25 Synthesis of the New DNA Strands The Lagging Strand is discontinuouslyThe Lagging Strand is synthesized discontinuously against overall direction of replication This strand is made in MANY short segments It is replicated from the replication fork toward the origin RNA Primer Leading Strand DNA Polymerase 5’5’ 5’ 3’ Lagging Strand 5’ 3’

26 Lagging Strand Segments Okazaki Fragments - lagging strandOkazaki Fragments - series of short segments on the lagging strand Must be joined together by an enzymeMust be joined together by an enzyme Lagging Strand RNAPrimerDNAPolymerase 3’ 5’ Okazaki Fragment

27 Joining of Okazaki Fragments The enzyme Ligase joins the Okazaki fragments together to make one strandThe enzyme Ligase joins the Okazaki fragments together to make one strand Lagging Strand Okazaki Fragment 2 DNA ligase DNA ligase Okazaki Fragment 1 5’ 3’

28 Replication of Strands Replication Fork Point of Origin

29 Proofreading New DNA DNA polymerase initially makes about 1 in 10,000 base pairing errorsDNA polymerase initially makes about 1 in 10,000 base pairing errors Enzymes proofread and correct these mistakesEnzymes proofread and correct these mistakes The new error rate for DNA that has been proofread is 1 in 1 billion base pairing errorsThe new error rate for DNA that has been proofread is 1 in 1 billion base pairing errors

30 Semiconservative Model of Replication Idea presented by Watson & CrickIdea presented by Watson & Crick TheThe two strands of the parental molecule separate, and each acts as a template for a new complementary strand New DNA consists of 1 PARENTAL (original) and 1 NEW strand of DNA Parental DNA DNA Template New DNA

31 DNA Damage & Repair Chemicals & ultraviolet radiation damage the DNA in our body cells Cells must continuously repair DAMAGED DNA Excision repair occurs when any of over 50 repair enzymes remove damaged parts of DNA DNA polymerase and DNA ligase replace and bond the new nucleotides together

32 Question: What would be the complementary DNA strand for the following DNA sequence? DNA 5’-CGTATG-3’

33 Answer: DNA 5’-CGTATG-3’ DNA 3’-GCATAC-5’