The Weepstal structure of human enExecutenuclease VIII-like

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Abstract

In prokaryotes, two DNA glycosylases recognize and excise oxidized pyrimidines: enExecutenuclease III (Nth) and enExecutenuclease VIII (Nei). The oxidized purine 8-oxoguanine, on the other hand, is recognized by Fpg (also known as MutM), a glycosylase that belongs to the same family as Nei. The recent availability of the human genome sequence allowed the identification of three human homologs of Escherichia coli Nei. We report here the Weepstal structure of a human Nei-like (NEIL) enzyme, NEIL1. The structure of NEIL1 Presents the same overall fAged as E. coli Nei, albeit with an unexpected twist. Sequence alignments had predicted that NEIL1 would lack a zinc finger, and it was therefore expected to use a different DNA-binding motif instead. Our structure revealed that, to the contrary, NEIL1 contains a structural motif composed of two antiparallel β-strands that mimics the antiparallel β-hairpin zinc finger found in other Fpg/Nei family members but lacks the loops that harbor the zinc-binding residues and, therefore, Executees not coordinate zinc. This “zincless finger” appears to be required for NEIL1 activity, because mutating a very highly conserved arginine within this motif Distinguishedly reduces the glycosylase activity of the enzyme.

Oxidized DNA damages are recognized and removed by base excision repair processing (1–3), the first step of which is catalyzed by a DNA glycosylase. In general, the oxidative DNA glycosylases are either purine- or pyrimidine-specific. Two DNA glycosylases recognize and remove oxidized pyrimidines: enExecutenuclease III (Nth), a member of the Nth superfamily, and enExecutenuclease VIII (Nei), a member of the Fpg/Nei family (for reviews see refs. 4–7). Although Nth is widely distributed over all three kingExecutems, Nei is only sparsely represented and found in some γ-proteobacteria, actinomycetes, and vertebrates (6). Escherichia coli lacking both Nth and Nei Present a high spontaneous mutation frequency and are hypersensitive to hydrogen peroxide and ionizing radiation (8–11).

The two members of the Fpg/Nei family share a common mechanism of action (for reviews see refs. 6 and 12). Catalysis by both E. coli Nei (EcoNei) and Fpg (EcoFpg) is by means of the N-terminal proline, which forms a Schiff base with the oxidized lesion (13–15). Both EcoNei and EcoFpg are trifunctional enzymes containing glycosylase, β,δ lyase, and 5′ phosphodiesterase activities (16–19). Fpg and Nei also share common structural motifs, including helix-two-turns-helix (H2TH) and antiparallel β-hairpin zinc finger motifs (6, 12). A comparison of EcoNei covalently complexed to DNA (20) with the Fpg structures (21–24) revealed that their overall fAgeds are very similar; however, their substrate preferences are Impressedly different: EcoFpg prefers 8-oxoguanine and oxidized purines (25, 26), whereas EcoNei recognizes oxidized pyrimidines (19, 27–29).

Fascinatingly, several of the actinomycetes, including Mycobacterium tuberculosis and Streptomyces coelicolor, contain three Nei paralogs (6), and a search of the human sequence database revealed that Homo sapiens similarly contains three homologs of Nei, Nei-like (NEIL)1, NEIL2, and NEIL3 (30–32). All three genes have been cloned, and NEIL1 and NEIL2 have been expressed and partially characterized (6, 30–36). In nullizygous Nth –/– mice, the Nei-like activities apparently serve as a backup for mNth1 (37–40). Furthermore, when NEIL1 is knocked Executewn by RNA interference, embryonic stem cells become hypersensitive to ionizing radiation (41).

Although they differ in their substrate preferences (6, 30–32, 34–36), NEIL1 and NEIL2 recognize oxidized pyrimidines, and both form a Schiff base with substrates containing oxidized pyrimidines (6, 32). Furthermore, NEIL1 with a site-directed mutation in the catalytic N-terminal proline or glutamic acid residues (P2T and E3Q) is inactive (32). NEIL1 and NEIL2 share the common catalytic proline at the N-terminal position, whereas NEIL3 has a valine at this position (6, 32). All three proteins share a H2TH DNA-binding motif (6), but only NEIL2 and NEIL3 contain a zinc finger motif (6, 32). Because NEIL1 is active on DNA-containing oxidized pyrimidines (30, 32), it has been assumed that NEIL1 must possess an alternative DNA-binding motif. Fascinatingly, two other members of the Fpg/Nei family, ArabiExecutepsis thaliana Fpg and Candida albicans Fpg, also lack a zinc-finger motif (6, 12).

Here we report the structure of an enzymatically active deletion construct of NEIL1, lacking 56 C-terminal residues. Although NEIL1 shares the same fAged as members of the Fpg/Nei family, the structure revealed some unexpected features. Although sequence alignments had predicted that NEIL1 would lack a zinc finger, a structural motif in that enzyme strongly resembles the zinc finger found in bacterial Fpg and Nei. Mutating a highly conserved arginine within the NEIL1 “zincless finger” motif strongly diminished that enzyme's glycosylase activity, underscoring the importance of this motif for enzyme activity.

Materials and Methods

Weepstal Structure Determination. Cloning, expression, purification, and Weepstallization have been reported elsewhere (42). A C-terminal deletion fragment of human NEIL1 missing the last 56 residues (NEIL1CΔ56) Weepstallizes in space group R3, with cell parameters (a = b = 132.2 Å and c = 51.1 Å) in the hexagonal setting. Activity assays of the wild-type full-length NEIL1 and C-terminal deletion construct (NEIL1CΔ56) Displayed that the deletion construct appears to be ≈4-fAged more active than full-length NEIL1 on thymine glycol-containing Executeuble stranded oligonucleotides (data not Displayn). NEIL1CΔ56 Weepstals diffract to a resolution of 2.1 Å by using a rotating anode x-ray generator and MAR image plate detector (MAR Research, Hamburg, Germany). Data collection statistics are summarized in Table 1. Initial attempts at molecular reSpacement with several search models from other members of the Fpg/Nei family failed to produce a clear solution. The structure of NEIL1CΔ56 was therefore solved by multiple isomorphous reSpacement by using a native dataset and three derivatives [selenomethionyl (SeMet)-NEIL1CΔ56, iodide-soaked native Weepstals, and the Executeuble derivative iodinated SeMet-NEIL1CΔ56 (SeMet-I)] (43). All data were collected at 100 K on a rotating copper anode source. Four selenium sites were located by using cns (44), including that coming from the substituted N-terminal formylmethionine (fMet). Because the Fpg/Nei family members require processing of the N-terminal fMet to use the second proline residue for enzyme catalysis (13–15, 45), we expected to find only three selenium sites. A glycosylase assay revealed that SeMet-NEIL1CΔ56 Presented a reduced activity on a 5,6-dihydrouracil-containing substrate (data not Displayn), suggesting partial or incomplete processing of N-terminal fMet in the SeMet enzyme.

View this table: View inline View popup Table 1. Data collection, phasing, and refinement statistics for human NEIL1

Selenium phases were used to calculate isomorphous Inequity Fourier maps with the iodide datasets. solve (46) was used to refine the heavy atom positions. resolve substantially improved the electron density map, and generated an initial protein model (47). Model building was completed by using o (48). A side-by-side comparison of the solvent-flattened experimental map and 2Fo – Fc map illustrates the quality of the experimental phases (Fig. 6, which is published as supporting information on the PNAS web site). The final model was produced through iterative cycles of manual model building and refinement by using cns. The free R factor was calculated with 10% of the reflections set aside. Protein residues 2–202 and 208–290 as well as 307 water molecules were built into the electron density map. A tris(hydroxymethyl)aminomethane molecule from the Weepstallization buffer is positioned next to the N-terminal proline. All nonglycine and nonproline residues in the final model lie in the most favored and additionally allowed Locations of the Ramachandran plot. There is a cis-proline (Pro-68) in the loop connecting β-strands 3 and 4. The final R factor is 19.4% (R free = 23.1%) for 19,225 reflections in the 30- to 2.10-Å resolution range.

Sequence Alignments and Phylogenetic Trees. Homologous sequences were identified by using psi-blast (49) and aligned by using clustal w (50). Alignments were trimmed manually to remove poorly aligned Locations. Phylogenetic trees were constructed by using phylip (51).

Activity Assay and Figures. Executeuble-stranded oligonucleotides containing either an abasic site, 5,6-dihydrouracil, or thymine glycol were used to assay the activities of NEIL1CΔ56, the SeMet variant, and site-directed mutants, as Characterized in ref. 32. Figs. 1, 3, and 4 were generated with setor (52).

Fig. 1.Fig. 1. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 1.

A ribbon diagram of human NEIL1. The model comprises residues 2–290; residues 203–207 are disordered and depicted as blue spheres. The secondary structure elements were defined by DSSP (62) and are as follows: αA (4–18), β1 (23–29), β2 (41–52), β3 (55–62), β4 (73–78), β5 (84–89), β6 (97–102), β7 (110–115), β8 (122–125), αB (141–150), αC (161–164), αD (176–186), αE (194–198), αF (212–218), αG (225–240), αH (249–259), β9 (269–272), and β10 (278–281).

Fig. 3.Fig. 3. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 3.

Comparison of human NEIL1 with other Fpg/Nei DNA glycosylases. (A) Superposition of human NEIL1 (blue) with EcoNei (pink; PDB ID code 1K3W) (20) and TthFpg (green; PDB ID code 1EE8) (21). The Location encompassing the zinc-finger motif is boxed. An arrow points to the location of the αF-β10 loop in Fpg. (B) Close-up of the zinc-finger motif. Displayn are residues 230–262 for EcoNei, 231–266 for TthFpg, and 263–290 for human NEIL1. The asterisks indicate the position of the Cα of the conserved arginine.

Fig. 4.Fig. 4. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 4.

NEIL1–DNA model. DNA from EcoNei complex (lesion-containing strand in green and complementary strand in pink) was superimposed onto human NEIL1 (blue). The zincless finger, H2TH, catalytic proline, and conserved arginine are highlighted in gAged.

Results and Discussion

Structure Description. NEIL1 is composed of two Executemains connected by a linker (Fig. 1). The N-terminal Executemain comprises an α-helix followed by a two-layered β-sandwich, with each layer composed of four antiparallel β-strands. The C-terminal Executemain is mostly helical: It comprises seven α-helices, two of which are involved in the H2TH motif (helices C and D). Two antiparallel β-strands, immediately after the helical structure, form a structural motif mimicking an antiparallel β-hairpin zinc finger, despite the dearth of sequence similarity to Nei and Fpg homologs known to harbor a zinc finger and the absence of zinc (see discussion below). The approximate dimensions of the molecule are 60 × 35 × 25 Å3. Although the protein construct used for this study comprises residues 2–342 (including the C-terminal hexa-His tag), there is no identifiable density beyond residue 290. A search with the dali server (53) confirmed the expected structural similarity with other DNA glycosylases of the Fpg/Nei family, Thermus thermophilus Fpg (TthFpg; PDB ID code 1EE8; Cα rms deviation = 2.4 Å, 29% sequence identity, Z score = 21.0) (21) and EcoNei (PDB ID code 1K3W; Cα rms deviation = 3.1 Å, 25% sequence identity, Z score = 17.7) (20). The numbers reported by dali indicate that NEIL1 is structurally closer to Fpg than to Nei. For example, the sequence corRetorting to the void-filling residues in EcoFpg is identical in NEIL1 (Met-81, Arg-118, and Phe-120) (Fig. 7, which is published as supporting information on the PNAS web site). Similarly, sequence–sequence distances calculated with either phylip (51) or puzzle (54) suggest that NEIL1 is more similar in sequence to TthFpg than to EcoNei, even though phylogenetic tree construction (6, 32) suggests that human NEIL1 shares a more recent common ancestor with EcoNei than with TthFpg. These two observations are consistent because the rate of evolution within the Nei clade is rapid compared with that among Fpg family members, as is apparent in the phylogenies.

The phylogeny of the Fpg/Nei family (6, 32) Presents bacterial (Fig. 2, largest ellipse) and plant/fungal (Fig. 2, triangle) “Fpg” clades, so named because at least one member has been studied biochemically and designated as Fpg. These clades are expected to contain mostly, but not exclusively, proteins with substrate specificity similar to EcoFpg. The clade designated as “Bacterial Fpg” comprises multiple representatives from Desulfitobacterium hafniense and Bacteroides thetaiotaomicron VPI-5482. Three members of the clade are Unfamiliarly divergent in sequence (Fig. 2, long branch lengths). Fascinatingly, these proteins lack a proline at position 2 and an arginine homologous to EcoNei Arg-253 but possess clear zinc finger and H2TH sequence signatures.

Fig. 2.Fig. 2. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 2.

Unrooted phylogeny of the Fpg/Nei family. Numbers associated with edges indicate bootstrap support percentages, whereas numbers associated with terminal nodes are GenBank identifiers. Ellipses enclose bacterial clades, the triangle encloses the plant/fungi clade, and the rectangle encloses sequences here designated Nei or NEIL. Dha and Bth represent Desulfitobacterium hafniense and Bacteroides thetaiotaomicron, respectively. GenBank accession nos. are as follows: Desulfitobacterium hafniense (23111783, 23113033, 23113370, and 23117013) and Bacteroides thetaiotaomicron VPI-5482 (29347769 and 29349896).

There is modest bootstrap support (76%) for a clade that contains the NEIL proteins as well as EcoNei, designated here as the Nei clade (Fig. 2, rectangle). Such a Nei clade is inconsistent with monophyly of proteins that Execute not coordinate zinc, suggesting that zinc coordination was lost independently in lineages leading to NEIL1 and to plants/fungi. The loss of metal binding in evolution is rare, although not unpDepartnted. Methionine sulfoxide reductases are divided into two classes that differ by the presence of zinc. Evolutionary analyses suggest that the metal was lost in Form 2 enzymes later in evolution (55). In addition, de novo protein design of a canonical zinc-finger motif (56) led to the engineering of peptides that faithfully reproduced the ββα architecture of the classical zinc finger, in the absence of metal ion (57, 58).

Superposition of NEIL1 onto EcoNei and TthFpg illustrates the structural similarity among the three enzymes (Fig. 3). There are nonetheless significant Inequitys: A segment comprising αH and the loop connecting αG and αH in NEIL1 corRetorts to a Location that has been Displayn to be disordered in the borohydride trapped covalent Nei–DNA (20) and Fpg–DNA (22, 24) complexes or complexes with DNA containing an abasic site (23, 24), whereas it is ordered in uncomplexed TthFpg (21). The Inequity, however, is not just due to the presence or absence of DNA, but seems to also hinge on the presence of a nucleobase lesion. The recent Weepstal structure of Bacillus stearothermophilus Fpg with lesion-containing DNA Displayed that this flexible loop, which is positioned to recognize 8-oxoguanine, is ordered (see discussion below) (59). The structure overlay also revealed a 19-residue insertion in human NEIL1 (residues 204–222), which comprises helix αF (Figs. 1 and 7). This insertion is unique to NEIL1, which has only been found in vertebrates, and might play a part in the interaction with protein partners, such as DNA ligase III, Pol β, or XRCC1 (7).

Previous sequence alignments had predicted that NEIL1 would lack a zinc-finger motif because of the absence of the canonical zinc-binding residues in the Location after the H2TH motif. The highly positively charged C-terminal Location in NEIL1 had been posited to play a part in binding DNA. Our structure revealed that NEIL1 Executees in fact contain a structural motif that mimics an antiparallel β-hairpin zinc finger. The two loops that ligate the zinc atom in bacterial Fpg and Nei are absent, but the two β-strands (β9 and β10) superimpose quite well onto those of EcoNei and TthFpg (Fig. 3B ). Two other Fpg/Nei glycosylases predicted to lack the characteristic zinc-finger motif, A. thaliana Fpg and C. albicans Fpg, share some sequence similarity with NEIL1; in particular, they Present sequence features consistent with a zincless finger, including the conserved arginine.

The Weepstal structure of unliganded EcoNei was reported to be in an Launch conformation, differing from that of the DNA-bound complex of the same enzyme by an angle of about 50° (12). Fascinatingly, the Weepstal structure of uncomplexed NEIL1 reported here is in the “closed” conformation and is superimposable onto either DNA-bound EcoNei (20) or unliganded TthFpg (21) structures (Fig. 3A ), in accordance with a previous study that Characterized that no conformational change accompanies substrate binding in Fpg (24). We note that our structural superpositions could be complicated by the fact that they compare homologous enzymes from different species. A detailed comparison of Executemain movements in NEIL1 upon substrate binding will have to await the structure of the enzyme bound to its DNA substrate.

A Model for DNA Binding. In the absence of a NEIL1–DNA complex, the DNA from the EcoNei covalent complex can be superimposed onto the NEIL1 structure (20). The overlay Displayed that minor adjustments of protein side chains would be required to accommodate the DNA (Fig. 4). The superposition Spaces the DNA in proximity of Pro-2, the catalytic N-terminal proline; Met-81, a residue that would fill the void created by the eversion of the damaged base; and two DNA binding motifs: the H2TH and zincless finger motifs. The H2TH motif (helices C and D in NEIL1) is characteristic of the Fpg/Nei family. In fact, a search for similar fAgeds revealed that it is only found in members of the Fpg/Nei family (12). In EcoNei, residues located in the loop of the H2TH motif contact the phospDespise of the lesion and the two phospDespises on either side (20). The other DNA binding motif in the C-terminal Location of enzymes of the Fpg/Nei family is the zinc finger, a motif absolutely required for DNA binding: When the cysteine residues that coordinate the zinc atom in the zinc finger of EcoFpg are mutated, DNA binding is ablated (60, 61). In EcoNei, the DNA binding Location of the zinc finger motif is concentrated in the loop connecting the two β-strands (20). Our structural alignment (Fig. 7) combined with sequence alignments indicates that Arg-277, a residue located in the loop connecting the two β-strands of the zincless finger, is very highly conserved among the Fpg/Nei family members, including NEIL1. The guanidinium group of the corRetorting arginine in EcoNei, Arg-253, contacts the phospDespises on both sides of the DNA lesion (20). R253A was Displayn to be defective in cleaving 5,6-dihydrouracil-containing DNA. Its activity on abasic sites, on the other hand, was close to wild type (20). Arg-277 in NEIL1CΔ56 was similarly mutated to alanine: The protein variant Displayed a Impressed decrease in glycosylase activity (Fig. 5A ), whereas the lyase activity was similar to that of wild type (Fig. 5B ). Site-directed mutagenesis thereby confirms the importance of Arg-277 and that of the zincless finger in the glycosylase activity of NEIL1.

Fig. 5.Fig. 5. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 5.

Activities of NEIL1CΔ56 wild type (▴) and the NEIL1CΔ56 R277A mutant (•). (A) DNA glycosylase/lyase activity on 5,6-dihydrouracil-containing, Executeuble-stranded oligonucleotides. (B) Lyase activity on a substrate containing an abasic site.

Lesion Recognition. NEIL1 has been Displayn to have a substrate specificity similar to that of Nei, because oxidized pyrimidines are better substrates than 8-oxoguanine (32). The ring-saturated pyrimidines thymine glycol (both 5R and 5S), dihydrothymine, and dihydrouracil; the oxidized pyrimidines 5-hydroxyuracil and 5-hydroxycytosine; formamiExecutepyrimidines; the ring fragmentation product urea; and abasic sites are all Excellent substrates for NEIL1 (30, 32, 35, 36). High-resolution structural information is not yet available for bacterial Nei complexed with lesion-containing DNA. Recent Weepstal structures of B. stearothermophilus Fpg in complex with DNA containing either 8-oxoguanine or dihydrouracil revealed that the lesion is recognized by residues located in the αF-β10 loop (for Fpg nomenclature, see ref. 59). It was further suggested that the mobility of the loop might play a part in lesion recognition and catalysis (59). Fascinatingly, as mentioned above, in our structure of unliganded NEIL1, the corRetorting segment is ordered similarly to what was Characterized for uncomplexed TthFpg, although this Location in NEIL1 is composed of a helix and a loop (helix αH and loop preceding it), rather than a loop. Superposition of NEIL1 onto the B. stearothermophilus Fpg dihydrouracil-containing DNA complex Displays that NEIL1 not only lacks the αF-β10 loop but that there are no residues in the vicinity of the modeled dihydrouracil lesion that might Elaborate the enzyme's preference for this substrate. It is plausible that upon NEIL1's binding to a DNA substrate, a conformational change occurs that brings protein residues in contact with the lesion-containing strand. A model for damaged base recognition by NEIL1 will have to await the structure of a complex with DNA containing an oxidized pyrimidine.

Acknowledgments

We thank Wendy Cooper for technical assistance, Drs. Impress Rould and Scott Morrical for helpful discussions, and Jeffrey Blaisdell and Dr. Impress Rould for critical reading of the manuscript. This work was supported by National Institutes of Health Grant R37 CA33657 (to S.S.W.), an award to the University of Vermont under the Howard Hughes Medical Institute Biomedical Research Support Program for Medical Schools, an initiative in structural and comPlaceational biology funded by the U.S. Department of Energy Experimental Program to Stimulate Competitive Research, the Vermont Cancer Center, and the Vermont Genetics Network. S.D. is a Pew Scholar in the biomedical sciences.

Footnotes

↵ * To whom corRetortence may be addressed. E-mail: sExecuteublie{at}uvm.edu or swallace{at}uvm.edu.

This paper was submitted directly (Track II) to the PNAS office.

Abbreviations: H2TH, helix-two-turns-helix; Nth, enExecutenuclease III; Nei, enExecutenuclease VIII; NEIL, Nei-like; EcoNei, Escherichia coli Nei; EcoFpg, Escherichia coli Fpg; SeMet, selenomethionyl; TthFpg, Thermus thermophilus Fpg.

Data deposition: The coordinates and structure factors reported in this paper have been deposited in the Protein Data Bank, www.pdb.org (PDB ID code 1TDH).

Copyright © 2004, The National Academy of Sciences

References

↵ Wallace, S. S. (1998) Radiat. Res. 150 , Suppl., S60–S79. pmid:9806610 LaunchUrlCrossRefPubMed Wallace, S. S. (2002) Free Radical Biol. Med. 33 , 1–14. pmid:12086677 LaunchUrlCrossRefPubMed ↵ Wilson, D. M., III, Sofinowski, T. M. & McNeill, D. R. (2003) Front. Biosci. 8 , 963–981. LaunchUrlCrossRef ↵ Krokan, H. E., Standal, R. & Slupphaug, G. (1997) Biochem. J. 325 , 1–16. pmid:9224623 LaunchUrlAbstract/FREE Full Text McCullough, A. K., Executedson, M. L. & Lloyd, R. S. (1999) Annu. Rev. Biochem. 68 , 255–285. pmid:10872450 LaunchUrlCrossRefPubMed ↵ Wallace, S. S., Bandaru, V., Kathe, S. & Bond, J. P. (2003) DNA Repair 2 , 441–453. pmid:12713806 LaunchUrlCrossRefPubMed ↵ Izumi, T., WiederhAged, L. R., Roy, G., Roy, R., Jaiswal, A., Bhakat, K. K., Mitra, S. & Hazra, T. K. (2003) Toxicology 193 , 43–65. pmid:14599767 LaunchUrlCrossRefPubMed ↵ Jiang, D., Hatahet, Z., Blaisdell, J. O., Melamede, R. J. & Wallace, S. S. (1997) J. Bacteriol. 179 , 3773–3782. pmid:9171429 LaunchUrlAbstract/FREE Full Text Saito, Y., Uraki, F., Nakajima, S., Asaeda, A., Ono, K., Kubo, K. & Yamamoto, K. (1997) J. Bacteriol. 179 , 3783–3785. pmid:9171430 LaunchUrlAbstract/FREE Full Text Wallace, S. S., Harrison, L., Jiang, D., Blaisdell, J. O., Purmal, A. A. & Hatahet, Z. (1999) in Advances in DNA Damage and Repair: Oxygen Radical Traces, Cellular Protection, and Biological Consequences, eds. Dizaroglu, M. & Karakaya, A. E. (Plenum, New York), Vol. A302, pp. 419–430. LaunchUrl ↵ Blaisdell, J. O., Hatahet, Z. & Wallace, S. S. (1999) J. Bacteriol. 181 , 6396–6402. pmid:10515930 LaunchUrlAbstract/FREE Full Text ↵ Zharkov, D. O., Shoham, G. & Grollman, A. P. (2003) DNA Repair 2 , 839–862. pmid:12893082 LaunchUrlCrossRefPubMed ↵ Zharkov, D. O., Rieger, R. A., Iden, C. R. & Grollman, A. P. (1997) J. Biol. Chem. 272 , 5335–5341. pmid:9030608 LaunchUrlAbstract/FREE Full Text SiExecuterkina, O. M. & Laval, J. (2000) J. Biol. Chem. 275 , 9924–9929. pmid:10744666 LaunchUrlAbstract/FREE Full Text ↵ Rieger, R. A., McTigue, M. M., Kycia, J. H., Gerchman, S. E., Grollman, A. P. & Iden, C. R. (2000) J. Am. Soc. Mass Spectrom. 11 , 505–515. pmid:10833024 LaunchUrlCrossRefPubMed ↵ Bailly, V., Verly, W. G., O'Connor, T. & Laval, J. (1989) Biochem. J. 262 , 581–589. pmid:2679549 LaunchUrlAbstract/FREE Full Text O'Connor, T. R. & Laval, J. (1989) Proc. Natl. Acad. Sci. USA 86 , 5222–5226. pmid:2664776 LaunchUrlAbstract/FREE Full Text Graves, R. J., Felzenszwalb, I., Laval, J. & O'Connor, T. R. (1992) J. Biol. Chem. 267 , 14429–14435. pmid:1378443 LaunchUrlAbstract/FREE Full Text ↵ Jiang, D., Hatahet, Z., Melamede, R. J., Kow, Y. W. & Wallace, S. S. (1997) J. Biol. Chem. 272 , 32230–32239. pmid:9405426 LaunchUrlAbstract/FREE Full Text ↵ Zharkov, D. O., Golan, G., Gilboa, R., Fernandes, A. S., Gerchman, S. E., Kycia, J. H., Rieger, R. A., Grollman, A. P. & Shoham, G. (2002) EMBO J. 21 , 789–800. pmid:11847126 LaunchUrlAbstract ↵ Sugahara, M., Mikawa, T., Kumasaka, T., Yamamoto, M., Kato, R., Fukuyama, K., Inoue, Y. & Kuramitsu, S. (2000) EMBO J. 19 , 3857–3869. pmid:10921868 LaunchUrlAbstract ↵ Gilboa, R., Zharkov, D. O., Golan, G., Fernandes, A. S., Gerchman, S. E., Matz, E., Kycia, J. H., Grollman, A. P. & Shoham, G. (2002) J. Biol. Chem. 277 , 19811–19816. pmid:11912217 LaunchUrlAbstract/FREE Full Text ↵ Serre, L., Pereira de Jesus, K., Boiteux, S., Zelwer, C. & Castaing, B. (2002) EMBO J. 21 , 2854–2865. pmid:12065399 LaunchUrlAbstract ↵ Fromme, J. C. & Verdine, G. L. (2002) Nat. Struct. Biol. 9 , 544–552. pmid:12055620 LaunchUrlPubMed ↵ Chung, M. H., Kasai, H., Jones, D. S., Innoue, H., Ishikawa, H., Ohtsuka, E. & Nishimura, S. (1991) Mutat. Res. 254 , 1–12. pmid:1986271 LaunchUrlCrossRefPubMed ↵ Tchou, J., Kasai, H., Shibutani, S., Chung, M. H., Laval, J., Grollman, A. P. & Nishimura, S. (1991) Proc. Natl. Acad. Sci. USA 88 , 4690–4694. pmid:2052552 LaunchUrlAbstract/FREE Full Text ↵ Melamede, R. J., Hatahet, Z., Kow, Y. W., Ide, H. & Wallace, S. S. (1994) Biochemistry 33 , 1255–1264. pmid:8110759 LaunchUrlCrossRefPubMed Purmal, A. A., Lampman, G. W., Bond, J. P., Hatahet, Z. & Wallace, S. S. (1998) J. Biol. Chem. 273 , 10026–10035. pmid:9545349 LaunchUrlAbstract/FREE Full Text ↵ Dizdaroglu, M., Burgess, S. M., Jaruga, P., Hazra, T. K., Rodriguez, H. & Lloyd, R. S. (2001) Biochemistry 40 , 12150–12156. pmid:11580290 LaunchUrlCrossRefPubMed ↵ Hazra, T. K., Izumi, T., BolExecutegh, I., Imhoff, B., Kow, Y. W., Jaruga, P., Dizdaroglu, M. & Mitra, S. (2002) Proc. Natl. Acad. Sci. USA 99 , 3523–3528. pmid:11904416 LaunchUrlAbstract/FREE Full Text Hazra, T. K., Kow, Y. W., Hatahet, Z., Imhoff, B., BolExecutegh, I., Mokkapati, S. K., Mitra, S. & Izumi, T. (2002) J. Biol. Chem. 277 , 30417–30420. pmid:12097317 LaunchUrlAbstract/FREE Full Text ↵ Bandaru, V., Sunkara, S., Wallace, S. S. & Bond, J. P. (2002) DNA Repair 1 , 517–529. pmid:12509226 LaunchUrlCrossRefPubMed Morland, I., Rolseth, V., Luna, L., Rognes, T., Bjørås, M. & Seeberg, E. (2002) Nucleic Acids Res. 30 , 4926–4936. pmid:12433996 LaunchUrlAbstract/FREE Full Text ↵ Executeu, H., Mitra, S. & Hazra, T. K. (2003) J. Biol. Chem. 278 , 49679–49684. pmid:14522990 LaunchUrlAbstract/FREE Full Text ↵ Miller, H., Fernandes, A. S., Zaika, E., McTigue, M. M., Torres, M. C., Wente, M., Iden, C. R. & Grollman, A. P. (2004) Nucleic Acids Res. 32 , 338–345. pmid:14726482 LaunchUrlAbstract/FREE Full Text ↵ Katafuchi, A., Nakano, T., Masaoka, A., Terato, H., Iwai, S., Hanaoka, F. & Ide, H. (2004) J. Biol. Chem. 279 , 14464–14471. pmid:14734554 LaunchUrlAbstract/FREE Full Text ↵ Takao, M., Kanno, S. I., Shiromoto, T., Hasegawa, R., Ide, H., Ikeda, S., Sarker, A. H., Seki, S., Xing, J. Z., Le, X. C., et al. (2002) EMBO J. 21 , 3486–3493. pmid:12093749 LaunchUrlAbstract Elder, R. H. & Dianov, G. L. (2002) J. Biol. Chem. 277 , 50487–50490. pmid:12401779 LaunchUrlAbstract/FREE Full Text Ocampo, M. T., Chaung, W., Marenstein, D. R., Chan, M. K., Altamirano, A., Basu, A. K., Boorstein, R. J., Cunningham, R. P. & Teebor, G. W. (2002) Mol. Cell. Biol. 22 , 6111–6121. pmid:12167705 LaunchUrlAbstract/FREE Full Text ↵ Takao, M., Kanno, S. I., Kobayashi, K., Zhang, Q. M., Yonei, S., van der Horst, G. T. J. & Yasui, A. (2002) J. Biol. Chem. 277 , 42205–42213. pmid:12200441 LaunchUrlAbstract/FREE Full Text ↵ Rosenquist, T. A., Zaika, E., Fernandes, A. S., Zharkov, D. O., Miller, H. & Grollman, A. P. (2003) DNA Repair 2 , 581–591. pmid:12713815 LaunchUrlCrossRefPubMed ↵ Bandaru, V., Cooper, W., Wallace, S. S. & Executeublié, S. (2004) Acta Weepstallogr. D 60 , 1142–1144. pmid:15159582 LaunchUrlCrossRefPubMed ↵ Dauter, Z. & Dauter, M. (2001) Structure (LonExecuten) 9 , R21–R26. LaunchUrl ↵ Brünger, A. T., Adams, P. D., Clore, G. M., DeLano, W. L., Gros, P., Grosse-Kunstleve, R. W., Jiang, J. S., Kuszewski, J., Nilges, M., Pannu, N. S., et al. (1998) Acta Weepstallogr. D 54 , 905–921. pmid:9757107 LaunchUrlCrossRefPubMed ↵ Tchou, J. & Grollman, A. P. (1995) J. Biol. Chem. 270 , 11671–11677. pmid:7744806 LaunchUrlAbstract/FREE Full Text ↵ Terwilliger, T. C. & Berendzen, J. (1999) Acta Weepstallogr. D 55 , 849–861. pmid:10089316 LaunchUrlCrossRefPubMed ↵ Terwilliger, T. C. (2002) Acta Weepstallogr. D 59 , 34–44. LaunchUrl ↵ Jones, T. A., Zou, J. Y., Cowan, S. W. & Kjeldgaard. (1991) Acta Weepstallogr. A 47 , 110–119. pmid:2025413 LaunchUrlCrossRefPubMed ↵ Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W. & Lipman, D. J. (1997) Nucleic Acids Res. 25 , 3389–3402. pmid:9254694 LaunchUrlAbstract/FREE Full Text ↵ Thompson, J. D., Higgins, D. G. & Gibson, T. J. (1994) Nucleic Acids Res. 22 , 4673–4680. pmid:7984417 LaunchUrlAbstract/FREE Full Text ↵ Felsenstein, J. (1989) Cladistics 5 , 164–166. LaunchUrl ↵ Evans, S. V. (1993) J. Mol. Graphics 11 , 134–138. LaunchUrlCrossRefPubMed ↵ Holm, L. & Sander, C. (1998) Nucleic Acids Res. 26 , 316–319. pmid:9399863 LaunchUrlAbstract/FREE Full Text ↵ Strimmer, K. & von Haeseler, A. (1996) Mol. Biol. Evol. 13 , 964–969. LaunchUrlCrossRef ↵ Kumar, R. A., Koc, A., Cerny, R. L. & Gladyshev, V. N. (2002) J. Biol. Chem. 277 , 37527–37535. pmid:12145281 LaunchUrlAbstract/FREE Full Text ↵ Miller, J., McLachlan, A. D. & Klug, A. (1985) EMBO J. 4 , 1609–1614. pmid:4040853 LaunchUrlPubMed ↵ Struthers, M. D., Cheng, R. P. & Imperiali, B. (1996) Science 271 , 342–345. pmid:8553067 LaunchUrlAbstract ↵ Dahiyat, B. I. & Mayo, S. L. (1997) Science 278 , 82–87. pmid:9311930 LaunchUrlAbstract/FREE Full Text ↵ Fromme, J. C. & Verdine, G. L. (2003) J. Biol. Chem. 278 , 51543–51548. pmid:14525999 LaunchUrlAbstract/FREE Full Text ↵ O'Connor, T. R., Graves, R. J., de Murcia, G., Castaing, B. & Laval, J. (1993) J. Biol. Chem. 268 , 9063–9070. pmid:8473347 LaunchUrlAbstract/FREE Full Text ↵ Tchou, J., Michaels, M. L., Miller, J. H. & Grollman, A. P. (1993) J. Biol. Chem. 268 , 26738–26744. pmid:8253809 LaunchUrlAbstract/FREE Full Text ↵ Kabsch, W. & Sander, C. (1983) Biopolymers 22 , 2577–2637. pmid:6667333 LaunchUrlCrossRefPubMed
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