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Titolo:
AP site structural determinants for Fpg specific recognition
Autore:
Castaing, B; Fourrey, JL; Hervouet, N; Thomas, M; Boiteux, S; Zelwer, C;
Indirizzi:
Univ Orleans, CNRS, UPR 4301, Ctr Biophys Mol, F-45071 Orleans 2, France Univ Orleans Orleans France 2 Ctr Biophys Mol, F-45071 Orleans 2, France CNRS, Inst Chim Subst Nat, UPR 2301, F-91198 Gif Sur Yvette, France CNRS Gif Sur Yvette France F-91198 2301, F-91198 Gif Sur Yvette, France Ctrses,d Nucl, Lab Radiobiol DNA, CEA, CNRS,UMR217, F-92265 Fontenay Aux Ro Ctr Etud Nucl Fontenay Aux Roses France F-92265 F-92265 Fontenay Aux Ro
Titolo Testata:
NUCLEIC ACIDS RESEARCH
fascicolo: 2, volume: 27, anno: 1999,
pagine: 608 - 615
SICI:
0305-1048(19990115)27:2<608:ASSDFF>2.0.ZU;2-I
Fonte:
ISI
Lingua:
ENG
Soggetto:
FORMAMIDOPYRIMIDINE-DNA GLYCOSYLASE; T4 ENDONUCLEASE-V; APURINIC APYRIMIDINIC SITES; ESCHERICHIA-COLI; ABASIC SITE; SUBSTRATE-SPECIFICITY; DAMAGED DNA; SACCHAROMYCES-CEREVISIAE; CATALYTIC MECHANISM; DELTA-ELIMINATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
55
Recensione:
Indirizzi per estratti:
Indirizzo: Castaing, B UnivOrleanss, CNRS, UPR 4301, Ctr Biophys Mol, Rue Charles Sadron, F-45071 Univ Orleans Rue Charles Sadron Orleans France 2 ron, F-45071
Citazione:
B. Castaing et al., "AP site structural determinants for Fpg specific recognition", NUCL ACID R, 27(2), 1999, pp. 608-615

Abstract

The binding of Escherichia coli and Lactococcus lactis Fapy-DNA glyosylase(Fpg) proteins to DNA containing either cyclic or non-cyclic abasic (AP) site analogs was investigated by electrophoretic mobility shift assay (EMSA)and by footprinting experiments. We showed that the reduced AP site is thebest substrate analog for the E. coli and L. lactis enzymes (K-Dapp = 0.26and 0.5 nM, respectively) as compared with the other analogs tested in this study (K-Dapp > 2.8 nM), The 1,3-propanediol (Pr) residue-containing DNA seems to be the minimal AP site structure allowing a Fpg specific DNA binding, since the ethyleneglycol residue is not specifically bound by these enzymes. The newly described cyclopentanol residue is better recognized than tetrahydrofuran (for the E.coli Fpg, K-Dapp = 2.9 and 25 nM, respectively), These results suggest that the hemiacetal form of the AP site is negativelydiscriminated by the Fpg protein suggesting a hydrogen bond between the C4'-hydroxyl group of the sugar and a Fpg residue. High-resolution hydroxyl radical footprinting using a duplex containing Pr shows that Fpg binds to six nucleotides on the strand containing the AP site and only the base opposite the lesion on the undamaged complementary strand, This comparative studyprovides new information about the molecular mechanism involved in the FpgAP lyase activity.

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Documento generato il 05/12/20 alle ore 01:52:57