Structural insights into the first incision reaction during nucleotide excision repair.
Nucleotide excision repair is a highly conserved DNA repair mechanism present in all kingdoms of life. The incision reaction is a critical step for damage removal and is accomplished by the UvrC protein in eubacteria. No structural information is so far available for the 3' incision reaction. Here we report the crystal structure of the N-terminal catalytic domain of UvrC at 1.5 A resolution, which catalyzes the 3' incision reaction and shares homology with the catalytic domain of the GIY-YIG family of intron-encoded homing endonucleases. The structure reveals a patch of highly conserved residues surrounding a catalytic magnesium-water cluster, suggesting that the metal binding site is an essential feature of UvrC and all GIY-YIG endonuclease domains. Structural and biochemical data strongly suggest that the N-terminal endonuclease domain of UvrC utilizes a novel one-metal mechanism to cleave the phosphodiester bond.
Truglio
James J
JJ
Department of Pharmacological Sciences, State University of New York at Stony Brook, Stony Brook, NY, USA.
Rhau
Benjamin
B
Croteau
Deborah L
DL
Wang
Liqun
L
Skorvaga
Milan
M
Karakas
Erkan
E
DellaVecchia
Matthew J
MJ
Wang
Hong
H
Van Houten
Bennett
B
Kisker
Caroline
C
D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D020134:Catalytic Domain / Q000235:genetics; D002413:Cations, Divalent / Q000378:metabolism; D017124:Conserved Sequence; D018360:Crystallography, X-Ray; D004260:DNA Repair / Q000502:physiology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D055672:Static Electricity
D002413:Cations, Divalent; D004269:DNA, Bacterial; D029968:Escherichia coli Proteins; D011994:Recombinant Proteins; D004706:Endodeoxyribonucleases; C065074:I-TEVI endonuclease; C066567:UvrC protein, E coli