title: Mismatch repair proteins collaborate with methyltransferases in the repair of O(6)-methylguanine.
abstract: DNA repair is essential for combatting the adverse effects of damage to the genome. One example of base damage is O(6)-methylguanine (O(6)mG), which stably pairs with thymine during replication and thereby creates a promutagenic O(6)mG:T mismatch. This mismatch has also been linked with cellular toxicity. Therefore, in the absence of repair, O(6)mG:T mismatches can lead to cell death or result in G:C-->A:T transition mutations upon the next round of replication. Cysteine thiolate residues on the Ada and Ogt methyltransferase (MTase) proteins directly reverse the O(6)mG base damage to yield guanine. When a cytosine is opposite the lesion, MTase repair restores a normal G:C pairing. However, if replication past the lesion has produced an O(6)mG:T mismatch, MTase conversion to a G:T mispair must still undergo correction to avoid mutation. Two mismatch repair pathways in E. coli that convert G:T mispairs to native G:C pairings are methyl-directed mismatch repair (MMR) and very short patch repair (VSPR). This work examined the possible roles that proteins in these pathways play in coordination with the canonical MTase repair of O(6)mG:T mismatches. The possibility of this repair network was analyzed by probing the efficiency of MTase repair of a single O(6)mG residue in cells deficient in individual mismatch repair proteins (Dam, MutH, MutS, MutL, or Vsr). We found that MTase repair in cells deficient in Dam or MutH showed wild-type levels of MTase repair. In contrast, cells lacking any of the VSPR proteins MutS, MutL, or Vsr showed a decrease in repair of O(6)mG by the Ada and Ogt MTases. Evidence is presented that the VSPR pathway positively influences MTase repair of O(6)mG:T mismatches, and assists the efficiency of restoring these mismatches to native G:C base pairs.
authors.affiliation: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
authors.forename: Peter T
authors.initials: PT
authors.lastname: Rye
authors.forename: James C
authors.initials: JC
authors.lastname: Delaney
authors.forename: Chawita
authors.initials: C
authors.lastname: Netirojjanakul
authors.forename: Dana X
authors.initials: DX
authors.lastname: Sun
authors.forename: Jenny Z
authors.initials: JZ
authors.lastname: Liu
authors.forename: John M
authors.initials: JM
authors.lastname: Essigmann
mesh_terms: D053843:DNA Mismatch Repair; D004268:DNA-Binding Proteins / Q000378:metabolism*; D018274:Electroporation; D004926:Escherichia coli; D029968:Escherichia coli Proteins / Q000235:genetics; D006147:Guanine / Q000031:analogs & derivatives* / Q000378:metabolism; D008780:Methyltransferases / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific) / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D009841:Oligonucleotides; D006147:Guanine; C008449:O-(6)-methylguanine; D008780:Methyltransferases; D015265:Site-Specific DNA-Methyltransferase (Adenine-Specific); C499294:dam protein, E coli