1 / 54

Maintenance and expression of genetic information

Maintenance and expression of genetic information. Central Dogma: DNA RNA Protein. GAATTGCGCCTTTTG. 5’- GAATTGCGCCTTTTG -3 3’- CTTAACGCGGAAAAC -5’. Minor Groove. Major Groove. DNA can be supercoiled. Semi-conservative Replication of DNA

mayes
Download Presentation

Maintenance and expression of genetic information

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript


  1. Maintenance and expression of genetic information Central Dogma: DNA RNA Protein

  2. GAATTGCGCCTTTTG

  3. 5’-GAATTGCGCCTTTTG-3 3’-CTTAACGCGGAAAAC-5’

  4. Minor Groove Major Groove

  5. DNA can be supercoiled

  6. Semi-conservative Replication of DNA The Watson-Crick Model

  7. Proof of the Watson-Crick Model: The Meselson-Stahl Experiment

  8. # generations The Meselson-Stahl Experiment 0 0.3 0.7 1 1.1 1.5 1.9 2.3 3 4.1 0 and 1.9 mixed 0 and 4.1 mixed

  9. The Meselson-Stahl Experiment Starting DNA Heavy/Heavy 1st generation All Heavy/Light 2nd generation Two Heavy/Light Two Light/Light 3rd generation Two Heavy/Light Six Light/Light

  10. DNA Polymerase

  11. A 3’ hydroxyl group is necessary for addition of nucleotides

  12. DNA chain growth is driven by PPi release/hydrolysis 5’ 5’ 4’ 4’ 3’ 3’ 1’ 2’ 2’ 5’ 5’ 4’ 4’ 3’ 3’ 1’ 1’ 2’ 2’

  13. Accuracy of DNA polymerases is essential. --Error rate is less than 1 in 108 --Due in part to “reading” of complementary bases --also contains its own proofreading activity

  14. DNA Polymerase contains a Proofreading subunit

  15. Proofreading by DNA polymerase

  16. Proofreading by DNA polymerase

  17. Both Template strands are copied at a Replication Fork

  18. The polarity of DNA synthesis creates an asymmetry between the leading strand and the lagging strand at the replication fork

  19. Topoisomerase Protein complexes of the replication fork

  20. Protein complexes of the replication fork: DNA polymerase DNA primase DNA Helicase ssDNA binding protein Sliding Clamp Clamp Loader DNA Ligase DNA Topoisomerase

  21. DNA primase synthesizes an RNA primer to initiate DNA synthesis on the lagging strand

  22. Replication of the Lagging Strand

  23. DNA ligase seals nicks left by lagging strand replication

  24. DNA helicase unwinds the DNA duplex ahead of DNA polymerase creating single stranded DNA that can be used as a template

  25. DNA helicase moves along one strand of the DNA

  26. ssDNA binding proteins are required to “iron out” the unwound DNA

  27. ssDNA binding proteins bind to the sugar phosphate backbone leaving the bases exposed for DNA polymerase

  28. DNA polymerase is not very processive (ie it falls off the DNA easily). A “sliding clamp” is required to keep DNA polymerase on and allow duplication of long stretches of DNA

  29. A “clamp loader:” complex is required to get the clamp onto the DNA

  30. Lagging strand synthesis

  31. Topoisomerase PCNA MCM proteins RPC

  32. Ahead of the replication fork the DNA becomes supercoiled

  33. The supercoiling ahead of the fork needs to be relieved or tension would build up (like coiling as spring) and block fork progression.

  34. Supercoiling is relieved by the action of Topoisomerases. Type I topoisomerases: Make nicks in one DNA strands Can relieve supercoiling Type II topoisomersases Make nicks in both DNA strands (double strand break) Can relieve supercoiling and untangle linked DNA helices Both types of enzyme form covalent intermediates with the DNA

  35. Topoisomerase I Action

  36. Topoisomerase I Action

  37. Topoisomerase II Action

  38. Topoisomerase II Action

  39. Topoisomerases as drug targets: Because dividing cells require greater topoisomerase activity due to increased DNA synthesis, topoisomerase inhibitors are used as chemotherapeutic agents. E.g. Camptothecin -- Topo I inhibitor Doxorubicin -- Topo II inhibitor These drugs act by stablilzing the DNA-Topoisomerase complex. Also, some antibiotics are inhibitors of the bacterial-specific toposisomerase DNA gyrase e.g. ciprofloxacin

  40. DNA is replicated during S phase of the Cell Cycle

  41. In S phase, DNA replication begins at origins of replication that are spread out across the chromosome

  42. Each origin of replicaton Initiates the formation of bidirectional replication forks

More Related