4 基因突变与修复.pptVIP

In all organisms, NER has the same steps: Damage recognition Binding of a protein complex at the damaged site Double incision of the damaged strand several nucleotides away from the damaged site, on both the 5 and 3 sides Removal of the damage-containing fragment from between the two nicks Filling in of the resulting gap by a DNA polymerase Ligation Nucleotide Excision Repair in E. coli 2 UvrA proteins form a complex with one UvrB protein in an ATP-dependent reaction The complex recognizes UV damage by the bend in the helix The UvrA proteins dissociate from the complex after ATP hydrolysis. This leaves UvrB bound across from the damage Nucleotide Excision Repair in E. coli Now UvrB can recruit UvrC protein to the complex UvrC activates UvrB to nick the DNA 4 nts 3’ from the pyrimidine dimer Then UvrB activates UvrC to nick the DNA 7 ntds 5’ from the pyrimidine dimer This leaves a fragment of DNA containing the damage that can now be removed Nucleotide Excision Repair in E. coli A helicase, UvrD, uses ATP hydrolysis to power the unwinding of the damaged DNA fragment. This reomoves UvrC The gap in the DNA is now filled in by DNAPI or II, reomving uvrB in the process Finally, DNA ligase seals the nick NER in Humans Critical for repairing UV-induced damage (because we don’t do direct repair) Principal is the same as in bacteria: But the proteins are different (20-30) Defects in NER proteins cause genetic disorders NER in HumansXeroderma pigmentosum (XP) Genetic disorder with symptoms: -extreme sensitivity to sunlight (by ~age 2), and 1000X higher risk of skin cancer (by ~age 8) Defect is in repair of UV damage Gene mapping identified 7 repair proteins (called XP proteins) XP-C and XP-A recognize pyrimidine dimers XP-B and XP-D have helicase activity XP-G and XP-F have nuclease activity NER in Humans 4.2.3 Excision repair Base excision repair (BER) - fixes abnormal bases (uracil, hypoxanthine, alkylated bases) Nucleotide excision repair (NER)