title: Atomic structure of the DNA repair [4Fe-4S] enzyme endonuclease III.
abstract: The crystal structure of the DNA repair enzyme endonuclease III, which recognizes and cleaves DNA at damaged bases, has been solved to 2.0 angstrom resolution with an R factor of 0.185. This iron-sulfur [4Fe-4S] enzyme is elongated and bilobal with a deep cleft separating two similarly sized domains: a novel, sequence-continuous, six-helix domain (residues 22 to 132) and a Greek-key, four-helix domain formed by the amino-terminal and three carboxyl-terminal helices (residues 1 to 21 and 133 to 211) together with the [4Fe-4S] cluster. The cluster is bound entirely within the carboxyl-terminal loop with a ligation pattern (Cys-X6-Cys-X2-Cys-X5-Cys) distinct from all other known [4Fe-4S] proteins. Sequence conservation and the positive electrostatic potential of conserved regions identify a surface suitable for binding duplex B-DNA across the long axis of the enzyme, matching a 46 angstrom length of protected DNA. The primary role of the [4Fe-4S] cluster appears to involve positioning conserved basic residues for interaction with the DNA phosphate backbone. The crystallographically identified inhibitor binding region, which recognizes the damaged base thymine glycol, is a seven-residue beta-hairpin (residues 113 to 119). Location and side chain orientation at the base of the inhibitor binding site implicate Glu112 in the N-glycosylase mechanism and Lys120 in the beta-elimination mechanism. Overall, the structure reveals an unusual fold and a new biological function for [4Fe-4S] clusters and provides a structural basis for studying recognition of damaged DNA and the N-glycosylase and apurinic/apyrimidinic-lyase mechanisms.
authors.affiliation: Department of Molecular Biology, Scripps Research Institute, La Jolla, CA 92037.
authors.forename: C F
authors.initials: CF
authors.lastname: Kuo
authors.forename: D E
authors.initials: DE
authors.lastname: McRee
authors.forename: C L
authors.initials: CL
authors.lastname: Fisher
authors.forename: S F
authors.initials: SF
authors.lastname: O'Handley
authors.forename: R P
authors.initials: RP
authors.lastname: Cunningham
authors.forename: J A
authors.initials: JA
authors.lastname: Tainer
mesh_terms: D001426:Bacterial Proteins / Q000648:ultrastructure; D001483:Base Sequence; D003461:Crystallography; D003545:Cysteine / Q000737:chemistry; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000648:ultrastructure*; D043224:Deoxyribonuclease (Pyrimidine Dimer); D004706:Endodeoxyribonucleases / Q000648:ultrastructure*; D007506:Iron-Sulfur Proteins / Q000648:ultrastructure*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D007506:Iron-Sulfur Proteins; D009838:Oligodeoxyribonucleotides; D004706:Endodeoxyribonucleases; D043224:Deoxyribonuclease (Pyrimidine Dimer); D003545:Cysteine