MB-5-repair-08例析.ppt

Damage Caused by Alkylation of Bases Alkylating agents like ethylmethane sulfonate (EMS) add alkyl groups to bases Some alkylation don’t change base-pairing, innocuous Others cause DNA replication to stall Cytotoxic Lead to mutations if cell attempts to replicate without damage repair Third type change base-pairing properties of a base, so are mutagenic 4) Damage Caused by Radiation Ultraviolet rays Comparatively low energy Cause a moderate type of damage Result in formation of pyrimidine dimers Gamma and x-rays Much more energetic Ionize molecules around the DNA Form highly reactive free radicals that attack DNA Alter bases Break strands 2) Excision Repair Percentage of DNA damage products that can be handled by direct reversal is small Most damage involves neither pyrimidine dimers nor O6-alkylguanine Another repair mechanism is required, excision repair is the process that removes most damaged nucleotides Damaged DNA is removed Replaced with fresh DNA Base and nucleotide excision repair are both used Base Excision Repair Base excision repair (BER) acts on subtle base damage Begins with DNA glycosylase Extrudes a base in a damaged base pair Clips out the damaged base Leaves an apurinic or apyrimidinic site that attracts DNA repair enzymes DNA repair enzymes Remove the remaining deoxyribose phosphate Replace it with a normal nucleotide Base Excision Repair in E. coli DNA polymerase I fills in missing nucleotide in BER Base is removed the AP site remains – apurinic or apyrimidinic AP endonuclease cuts or nicks DNA strand Phosphodiesterase removes the AP sugar phosphate Pol I performs repair synthesis Nucleotide Excision Repair Nucleotide excision repair typically handles bulky damage that distorts DNA double helix NER in E. coli begins when damaged DNA is clipped by an endonuclease on either side of the lesion, sites 12-13 nt apart Enzyme system catalyzing nucleotide excision repair is excinuclease Allows damaged DNA to be removed as part of resulting 12-13-ba