Repair of DNA methylphosphotriesters through a metalloactivated cysteine nucleophile

Science. 1993 Aug 27;261(5125):1164-7. doi: 10.1126/science.8395079.

Abstract

The Escherichia coli Ada protein repairs methylphosphotriesters in DNA by direct, irreversible methyl transfer to one of its own cysteines. Upon methyl transfer, Ada acquires the ability to bind specific DNA sequences and thereby to induce genes that confer resistance to methylating agents. The amino-terminal domain of Ada, which comprises the methylphosphotriester repair and sequence-specific DNA binding elements, contains a tightly bound zinc ion. Analysis of the zinc binding site by cadmium-113 nuclear magnetic resonance and site-directed mutagenesis revealed that zinc participates in the autocatalytic activation of the active site cysteine and may also function as a conformational switch.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Binding Sites
  • Cadmium
  • Cysteine / metabolism
  • DNA / metabolism*
  • DNA Repair*
  • Escherichia coli Proteins*
  • Isotopes
  • Magnetic Resonance Spectroscopy
  • Methylation
  • Mutagenesis, Site-Directed
  • O(6)-Methylguanine-DNA Methyltransferase
  • Protons
  • Transcription Factors
  • Zinc / chemistry
  • Zinc / metabolism*

Substances

  • Bacterial Proteins
  • Escherichia coli Proteins
  • Isotopes
  • Protons
  • Transcription Factors
  • Cadmium
  • DNA
  • Ada protein, E coli
  • O(6)-Methylguanine-DNA Methyltransferase
  • Zinc
  • Cysteine