From Proteopedia

Revision as of 05:45, 19 March 2010

Template:STRUCTURE 1yqr 8-Oxoguanine Glycosylase

Introduction

8-Oxogunanine glycosylase (hOGG1) is a DNA glycosylase, coded from the OGG1 gene in humans; however, many homologs exist in different organisms and this enzyme was originally discovered in yeast. It is responsible for removing genotoxic lesions caused by oxidative damage in the presence of reactive oxygen species (ROS). Glycosylases, in general, are key enzymes for base excision repair and therefore maintaining integrity of DNA across the evolutionary spectrum. Once a lesion is successfully excised, the transcription machinery of the cell can repair the DNA strand; however, if it is not repaired mutagenesis occurs possibly leading to cancer and other degenerative diseases ^[1].

Function ^[1]

hOGG1 repairs (8-oxoG, GO); this lesion is caused by oxidative attack by ROS on G. It is a particularly dangerous, and stable mutation because GO can Hoogsteen base-pair with adenine causing G:C to T:A tranversions. hOGG1 is able to cleave the N-glycosylic bond between the deoxyribose moiety and GO leaving an apurinic-apyrimdinic (AP) site. It also has is the intrinsic ability to cleave the 3’ phosphodiester of the AP site by β-elimination, acting as an AP lyase, and making it a bifunctional glycosylase. hOGG1 has greater affinity for GO when is complementary to C, and also has catalytic activity towards other lesions such as formamidopyrimidines.

Structure

hOGG1 belongs to a super family of DNA repair enzymes that share a conserved two-domain fold containing a DNA binding motif followed by a Glycine/Proline rich stretch and a invariant Aspartate^[1]. This motif is necessary for interacting with DNA to recognize and catalyze the substrate ^[2]. As well, OGG1 is associated with two calcium ions that help stabilize the deformed DNA back bone at the site of the extruded lesion^[2].

Mechanism

Only 50,000 hOGG1 molecules protect the entire 6,000,000,000 nuclear base-pairs in a diploid cell ^[3]. For this reason it is obvious that hOGG1 must have an efficient mechanism to catalyze GO and discriminate between GO and G even though they differ only at the C8 and N7 ^[4].

Recognition

hOGG1 is able to discriminate GO from G with the help of a single hydrogen bond between a ^[5]. Additional structural studies have indicated that the GO is extruded from the DNA helix and inserted deeply into a where residues lining the pocket can directly interact to excise the lesion^[4].

Catalysis

To excise GO, Lys 249 acts as a nucleophile attacking the C1' carbon in a SN1 reaction, when GO is inserted into the catalytic pocket^[5]. Then hOGG1 AP lyase uses a conserved lysine residue as a nucleophile to generate a covalently linked enzyme-DNA adduct, which undergoes a series of subsequent transformation resulting in DNA strand exscission on the 3’ side of the lesion^[5].

References

1. ↑ ^{1.0 1.1 1.2} Lu AL, Li X, Gu Y, Wright PM, Chang DY. Repair of oxidative DNA damage: mechanisms and functions. Cell Biochem Biophys. 2001;35(2):141-70. PMID:11892789 doi:10.1385/CBB:35:2:141
2. ↑ ^{2.0 2.1} Bruner SD, Norman DP, Verdine GL. Structural basis for recognition and repair of the endogenous mutagen 8-oxoguanine in DNA. Nature. 2000 Feb 24;403(6772):859-66. PMID:10706276 doi:10.1038/35002510
3. ↑ Cappelli E, Hazra T, Hill JW, Slupphaug G, Bogliolo M, Frosina G. Rates of base excision repair are not solely dependent on levels of initiating enzymes. Carcinogenesis. 2001 Mar;22(3):387-93. PMID:11238177
4. ↑ ^{4.0 4.1} Banerjee A, Yang W, Karplus M, Verdine GL. Structure of a repair enzyme interrogating undamaged DNA elucidates recognition of damaged DNA. Nature. 2005 Mar 31;434(7033):612-8. PMID:15800616 doi:10.1038/nature03458
5. ↑ ^{5.0 5.1 5.2} Lingaraju GM, Sartori AA, Kostrewa D, Prota AE, Jiricny J, Winkler FK. A DNA glycosylase from Pyrobaculum aerophilum with an 8-oxoguanine binding mode and a noncanonical helix-hairpin-helix structure. Structure. 2005 Jan;13(1):87-98. PMID:15642264 doi:10.1016/j.str.2004.10.011