2. DNA Replication AP Biology Unit 3

3. (1928) Griffith: Bacteria can be “transformed”/given new traits Image taken without permission from http://www.nature.com/scitable/topicpage/isolating-hereditary-material-frederick- griffith-oswald-avery-336

4. (1944) Avery- McCloud- McCarty: the transforming substance is DNA Image taken without permission from http://www.nature.com/scitable/topicpage/isolating-hereditary- material-frederick-griffith-oswald-avery-336

5. Hershey & Chase Experiment Bacteriophages are viruses that infect bacteria Bacteriophages consist of DNA and protein (capsid) What part of the bacteriophage holds the instructions of how to make more viruses? –DNA

6. (1952) Hershey & Chase: DNA is the genetic material

7. Structure of DNA (1953) Wilkins and Franklin: Used X-ray crystallography to create an image of DNA; Franklin: sugar & phosphates were the backbone of DNA Watson and Crick: Used the X-ray crystallograph to determine the structure of DNA and the base pairing (A-T, C-G) Image taken without permission from http://www.makingthemodernworld.org.uk/stories/defi ant_modernism/01.ST.02/?scene=5

8. DNA Structure (review) Double Helix (2 strands of DNA) Complementary strands pair up (A & T, C & G)– hydrogen bonds Strands are antiparallel (5’ and 3’ ends)

9. DNA Replication When DNA is copied during S Phase of Interphase Basic Concept = create a new strand by matching complementary nucleotides to an existing strand

10. DNA Replication Replication is semi-conservative (one strand is old, one strand new)

11. Origins of Replication Where DNA Replication starts Differs between organisms –Prokaryotes = 1 origin of replication –Eukaryotes = many different origins of replication

12. General Process of DNA Replication Step 1: Initiator Proteins bind to origins of replication to begin replication –Attracts other enzymes involved in replication process

13. General Process Step 2: –DNA Helicase separates the DNA helix –Topoisomerase (Gyrase) prevents “overwinding” of DNA by “nicking” the DNA then resealing it. –Single Stranded Binding Proteins (SSBs) prevent double helix from reforming temporarily

14. Question… What kind of bonds does Helicase break? –Hydrogen bonds How do you think SSBs prevent the DNA helix from reforming? –Prevent hydrogen bonds from reforming

15. General Process Step 3: Primase builds an RNA primer at the starting from the 5’ end of the new DNA strand –Uses the 3’ end of existing DNA strand –Why? Because the DNA Polymerase III (main DNA building enzyme) needs something to build off of 3’3’ 5’5’ 5’5’ Primase RNA Primer DNA Polymerase III

16. General Process Step 4: DNA Polymerase III builds the new strand of DNA in a 5’ to 3’ direction –What kinds bonds are being formed to make a new strand of DNA? –Covalent bonds

17. Question… What other enzyme builds similarly to DNA Polymerase III? RNA Polymerase (don’t mix it up with DNA Polymerase III!)

18. Problem… How can both strands of DNA be replicated in a 5’- 3’ direction at the same time they are antiparallel? Answer: leading and lagging strands

19. Leading and Lagging Strands Leading strand is synthesized continuously in the direction of replication (goes in the same direction as helicase) Lagging strand is synthesized in short fragments the opposite direction of replication (opposite direction as helicase)

20. Questions… How many primers does the leading strand need? –Only 1– to start replication How many primers does the lagging strand need? –Many – one for each Okazaki fragment

21. Lagging Strand

22. DNA Ligase DNA Ligase seals Okazaki fragments together –Forms covalent bonds between nucleotides to create a continuous strand of DNA

23. Finishing DNA Replication Problem #1: There are still RNA nucleotides in the DNA (primers) Solution = DNA Polymerase I cuts out the RNA nucleotides and replaces them with DNA

24. Finishing DNA Replication Problem #2: Okazaki fragments and bases replaced by DNA Polymerase I are not attached to rest of DNA Solution = DNA Ligase seals everything together

25. Putting It All Together Label the diagram on Page 71 with the following terms: -SSBs- DNA Ligase -Leading Strand- Lagging Strand -Helicase- Primase -DNA Polymerase III- Primer -DNA Polymerase I

26. SSBs DNA Polymerase III Primase Primer DNA Pol III Lagging Strand DNA Polymerase I Leading Strand DNA Ligase Helicase