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DNA Repair Supplement

DNA Repair Supplement. Read and digest please. Things that Damage DNA. Radiation - Cosmic Chemicals - in the environment DNA Replication Accidents Heat (spontaneous chemical alterations). Types of DNA Damage.

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DNA Repair Supplement

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  1. DNA Repair Supplement Read and digest please

  2. Things that Damage DNA • Radiation - Cosmic • Chemicals - in the environment • DNA Replication Accidents • Heat (spontaneous chemical alterations)

  3. Types of DNA Damage • All four of the bases in DNA (A, T, C, G) can be covalently modified at various positions. • Mismatches of the normal bases because of a failure of proofreading during DNA replication. • Breaks in the backbone. • Crosslinks Covalent linkages can be formed between bases

  4. Race to Repair • The repair mechanism are all operating independently in the cell • DNA damage may be repaired by which ever one detects that damage first. • Many components of the different repair pathways are common between them.

  5. Repairing Damaged Bases • Direct Reversal of Base Damage • Excision Repair • Base Excision Repair (BER) • Nucleotide Excision Repair (NER) • Mismatch Repair (MMR)

  6. 1.Direct Reversal of Base Damage • Non-humans - reverse defect by using light - photoreactivation • Humans - reversal of defects by special enzymes called glycosylases - that remove the mismatched T restoring the correct C. This is done without the need to break the DNA backbone (in contrast to the mechanisms of excision repair described next).

  7. 2. EXCISION REPAIRa) Base Excision Repair (BER) • 4 step process • 1.removal of the damaged base (estimated to occur some 20,000 times a day in each cell in our body!) by a DNA glycosylase. We have at least 8 genes encoding different DNA glycosylases each enzyme responsible for identifying and removing a specific kind of base damage. • 2.removal of its deoxyribose phosphate in the backbone, producing a gap. We have two genes encoding enzymes with this function.

  8. 2. EXCISION REPAIRa) Base Excision Repair (BER) • 4 step process • 3.replacement with the correct nucleotide. This relies on DNA polymerase beta, one of at least 11 DNA polymerases encoded by our genes. • 4.ligation of the break in the strand. Two enzymes are known that can do this; both require ATP to provide the needed energy.

  9. 2. EXCISION REPAIRb) Nucleotide Excision Repair (NER) • NER differs from BER in several ways. • It uses different enzymes. • Even though there may be only a single "bad" base to correct, its nucleotide is removed along with many other adjacent nucleotides • that is, NER removes a large "patch" around the damage.

  10. 2. EXCISION REPAIRb) Nucleotide Excision Repair (NER) • The damage is recognized by one or more protein factors that assemble at the location. • The DNA is unwound producing a "bubble". • Cuts are made on both the 3' side and the 5' side of the damaged area so the tract containing the damage can be removed. • A fresh burst of DNA synthesis - using the intact (opposite) strand as a template - fills in the correct nucleotides. The DNA polymerases responsible are designated polymerase delta and epsilon. • A DNA ligase covalent binds the fresh piece into the backbone.

  11. 2. EXCISION REPAIRc) Mismatch Repair (MMR) • Mismatch repair deals with correcting mismatches of the normal bases; that is, failures to maintain normal Watson-Crick base pairing (A･T, C･G)

  12. 2. EXCISION REPAIRc) Mismatch Repair (MMR) • It can enlist the aid of enzymes involved in both base-excision repair (BER) and nucleotide-excision repair (NER) as well as using enzymes specialized for this function • To repair mismatched bases, the system has to know which base is the correct one. • In E. coli, this is achieved by a special methylase called the "Dam methylase", which can methylate all adenines that occur within (5')GATC sequences. • Immediately after DNA replication, the template strand has been methylated, but the newly synthesized strand is not methylated yet. • Thus, the template strand and the new strand can be distinguished.

  13. 2. EXCISION REPAIRc) Mismatch Repair (MMR)

  14. 2. EXCISION REPAIRc) Mismatch Repair (MMR)

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