Nucleic Acids Research, 1993, Vol. 21, No. 12 2899-2905
© 1993
MOLECULAR BIOLOGY |
Cleavage and binding of a DNA fragment containing a single 8-oxoguanine by wild type and mutant FPG proteins
Laboratoire de Biologie Structurale du CNRS 1 av. de la Terrasse, 91198-Gif sur Yvette, France 1Sektion Polymere, Universität Ulm Albert Einstein Allee 11, W-7900 Ulm 2Lab. für Molekularbiologie and Allgemeine Pathologie, Universität Ulm Neuherbergstr. 11, W-8000 Munich 45, Germany 3LA147 CNRS, U140 INSERM, Institut Gustave Roussy, Groupe ‘Réparation des Lésions Radio- et Chimio-induites’ 94800-Villejuif, France
*To whom correspondence should be addressed
Received February 23, 1993. Revised May 27, 1993. Accepted May 27, 1993.
A 34-mer oligonucleotide containing a single 7,8-dihydro-8-oxoguanlne (8-OxoG) residue was used to study the enzymatic and DNA binding properties of the Fpg protein from E.coll. The highest rates of incision of the 8-OxoG containing strand by the Fpg protein were observed for duplexes where 8-OxoG was opposite C (*G/C) or T (*G/T). In contrast, the rates of incision of duplexes containing 8-OxoG opposite G (*G/G) and A (*G/A) were 5-fold and 200-fold slower. Gel retardation studies showed that the Fpg protein had a strong affinity for duplexes where the 8-OxoG was opposite pyrimidines and less affinity for duplexes where the 8-OxoG was opposite purines. Koapp values were 0.6 nM (*G/C), 1.0 nM (*G/T), 6.0 nM (*G/G) and 16.0 nM (*G/A). The Fpg protein also binds to unmodified (G/C) duplex and a KDapp of 90 nM was measured. The cleavage and binding of the (*G/C) duplex were also studied using bacterial crude lysates. Wild type E.coll crude extract incised the 8-OxoG containing strand and formed a specific retardation complex with the (*G/C) duplex. These two reactions were mediated by the Fpg protein, since they were not observed with a crude extract from a bacterial strain whose fpg gene was inactivated. Furthermore, we have studied the properties of 6 mutant Fpg proteins with Cys
Gly mutations. The results showed that the 2 Fpg proteins with CysGly mutations outside the zinc finger sequence cleaved the 8-OxoG containing strand, formed complexes with the (*G/C) duplex and suppressed the mutator phenotype of the fpg-1 mutant. In contrast, the 4 Fpg proteins with CysGly mutations within the zinc finger motif neither cleave nor bind the (*G/C) duplex, nor do these proteins suppress the fpg-1 mutator phenotype.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M. Pastoriza-Gallego, J. Armier, and A. Sarasin Transcription through 8-oxoguanine in DNA repair-proficient and Csb /Ogg1 DNA repair-deficient mouse embryonic fibroblasts is dependent upon promoter strength and sequence context Mutagenesis, September 1, 2007; 22(5): 343 - 351. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Rogacheva, A. Ishchenko, M. Saparbaev, S. Kuznetsova, and V. Ogryzko High Resolution Characterization of Formamidopyrimidine-DNA Glycosylase Interaction with Its Substrate by Chemical Cross-linking and Mass Spectrometry Using Substrate Analogs J. Biol. Chem., October 27, 2006; 281(43): 32353 - 32365. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. A. Ishchenko, X. Yang, D. Ramotar, and M. Saparbaev The 3'->5' Exonuclease of Apn1 Provides an Alternative Pathway To Repair 7,8-Dihydro-8-Oxodeoxyguanosine in Saccharomyces cerevisiae Mol. Cell. Biol., August 1, 2005; 25(15): 6380 - 6390. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. Coste, M. Ober, T. Carell, S. Boiteux, C. Zelwer, and B. Castaing Structural Basis for the Recognition of the FapydG Lesion (2,6-Diamino-4-hydroxy-5-formamidopyrimidine) by Formamidopyrimidine-DNA Glycosylase J. Biol. Chem., October 15, 2004; 279(42): 44074 - 44083. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. A. van der Kemp, J.-B. Charbonnier, M. Audebert, and S. Boiteux Catalytic and DNA-binding properties of the human Ogg1 DNA N-glycosylase/AP lyase: biochemical exploration of H270, Q315 and F319, three amino acids of the 8-oxoguanine-binding pocket Nucleic Acids Res., January 29, 2004; 32(2): 570 - 578. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Scaramozzino, G. Sanz, J. M. Crance, M. Saparbaev, R. Drillien, J. Laval, B. Kavli, and D. Garin Characterisation of the substrate specificity of homogeneous vaccinia virus uracil-DNA glycosylase Nucleic Acids Res., August 15, 2003; 31(16): 4950 - 4957. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Li and A-L. Lu Molecular Cloning and Functional Analysis of the MutY Homolog of Deinococcus radiodurans J. Bacteriol., November 1, 2001; 183(21): 6151 - 6158. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Kobune, Y. Xu, C. Baum, M. R. Kelley, and D. A. Williams Retrovirus-mediated Expression of the Base Excision Repair Proteins, Formamidopyrimidine DNA Glycosylase or Human Oxoguanine DNA Glycosylase, Protects Hematopoietic Cells from N,N',N''-Triethylenethiophosphoramide (thioTEPA)-induced Toxicity in Vitro and in Vivo Cancer Res., July 1, 2001; 61(13): 5116 - 5125. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Kamiya and H. Kasai 2-Hydroxy-dATP is incorporated opposite G by Escherichia coli DNA polymerase III resulting in high mutagenicity Nucleic Acids Res., April 1, 2000; 28(7): 1640 - 1646. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Saparbaev, J.-C. Mani, and J. Laval Interactions of the human, rat, Saccharomyces cerevisiae and Escherichia coli 3-methyladenine-DNA glycosylases with DNA containing dIMP residues Nucleic Acids Res., March 15, 2000; 28(6): 1332 - 1339. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Asagoshi, T. Yamada, H. Terato, Y. Ohyama, Y. Monden, T. Arai, S. Nishimura, H. Aburatani, T. Lindahl, and H. Ide Distinct Repair Activities of Human 7,8-Dihydro-8-oxoguanine DNA Glycosylase and Formamidopyrimidine DNA Glycosylase for Formamidopyrimidine and 7,8-Dihydro-8-oxoguanine J. Biol. Chem., February 18, 2000; 275(7): 4956 - 4964. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Gogos and N. D. Clarke Characterization of an 8-Oxoguanine DNA Glycosylase from Methanococcus jannaschii J. Biol. Chem., October 22, 1999; 274(43): 30447 - 30450. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Bauche and J. Laval Repair of Oxidized Bases in the Extremely Radiation-Resistant Bacterium Deinococcus radiodurans J. Bacteriol., January 1, 1999; 181(1): 262 - 269. [Abstract] [Full Text]
|
|
|
|
|
|
B. Sun, K. A. Latham, M. L. Dodson, and R. S. Lloyd Studies on the Catalytic Mechanism of Five DNA Glycosylases J. Biol. Chem., August 18, 1995; 270(33): 19501 - 19508. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. Castaing, C. Zelwer, J. Laval, and S. Boiteux HU Protein of Escherichia coli Binds Specifically to DNA That Contains Single-strand Breaks or Gaps J. Biol. Chem., April 28, 1995; 270(17): 10291 - 10296. [Abstract] [Full Text] [PDF]
|
|