TY - JOUR
T1 - Genome and cancer single nucleotide polymorphisms of the human NEIL1 DNA glycosylase
T2 - Activity, structure, and the effect of editing
AU - Prakash, Aishwarya
AU - Carroll, Brittany L.
AU - Sweasy, Joann B.
AU - Wallace, Susan S.
AU - Doublié, Sylvie
N1 - Funding Information:
We would like to thank Dr. Cynthia J. Burrows, University of Utah, for providing oligodeoxyribonucleotides containing spiroiminodihydantoin, and guanidinohydantoin; Dr. Carmelo Rizzo (Vanderbilt) for the oligodeoxynucleotide containing methyl-FapyG; Drs. Mark Rould, and Brian E. Eckenroth for helpful crystallographic suggestions; Drs. Jeff Bond and Julie Dragon for help with identifying the variants. This work was supported by National Institutes of Health Grant P01CA098993 awarded by the National Cancer Institute . AP was also supported by a J. Walter Juckett post-doctoral fellowship awarded by the Lake Champlain Cancer Research Organization and the Vermont Cancer Center . Support from the Vermont Cancer Center is acknowledged.
PY - 2014/2
Y1 - 2014/2
N2 - The repair of free-radical oxidative DNA damage is carried out by lesion-specific DNA glycosylases as the first step of the highly conserved base excision repair (BER) pathway. In humans, three orthologs of the prototypical endonuclease VIII (Nei), the Nei-like NEIL1-3 enzymes are involved in the repair of oxidized DNA lesions. In recent years, several genome and cancer single-nucleotide polymorphic variants of the NEIL1 glycosylase have been identified. In this study we characterized four variants of human NEIL1: S82C, G83D, P208S, and δE28, and tested their ability to excise pyrimidine-derived lesions such as thymine glycol (Tg), 5-hydroxyuracil (5-OHU), and dihydrouracil (DHU) and the purine-derived guanidinohydantoin (Gh), spiroiminodihydantoin 1 (Sp1), and methylated 2,6-diamino-4-hydroxy-5-formamidopyrimidine (MeFapyG). The P208S variant has near wild-type activity on all substrates tested. The S82C and δE28 variants exhibit decreased Tg excision compared to wild-type. G83D displays little to no activity with any of the substrates tested, with the exception of Gh and Sp1. Human NEIL1 is known to undergo editing whereby the lysine at position 242 is recoded into an arginine. The non-edited form of NEIL1 is more efficient at cleaving Tg than the R242 form, but the G83D variant does not cleave Tg regardless of the edited status of NEIL1. The corresponding G86D variant in Mimivirus Nei1 similarly lacks glycosylase activity. A structure of a G86D-DNA complex reveals a rearrangement in the β4/5 loop comprising Leu84, the highly-conserved void-filling residue, thereby providing a structural rationale for the decreased glycosylase activity of the glycine to aspartate variant.
AB - The repair of free-radical oxidative DNA damage is carried out by lesion-specific DNA glycosylases as the first step of the highly conserved base excision repair (BER) pathway. In humans, three orthologs of the prototypical endonuclease VIII (Nei), the Nei-like NEIL1-3 enzymes are involved in the repair of oxidized DNA lesions. In recent years, several genome and cancer single-nucleotide polymorphic variants of the NEIL1 glycosylase have been identified. In this study we characterized four variants of human NEIL1: S82C, G83D, P208S, and δE28, and tested their ability to excise pyrimidine-derived lesions such as thymine glycol (Tg), 5-hydroxyuracil (5-OHU), and dihydrouracil (DHU) and the purine-derived guanidinohydantoin (Gh), spiroiminodihydantoin 1 (Sp1), and methylated 2,6-diamino-4-hydroxy-5-formamidopyrimidine (MeFapyG). The P208S variant has near wild-type activity on all substrates tested. The S82C and δE28 variants exhibit decreased Tg excision compared to wild-type. G83D displays little to no activity with any of the substrates tested, with the exception of Gh and Sp1. Human NEIL1 is known to undergo editing whereby the lysine at position 242 is recoded into an arginine. The non-edited form of NEIL1 is more efficient at cleaving Tg than the R242 form, but the G83D variant does not cleave Tg regardless of the edited status of NEIL1. The corresponding G86D variant in Mimivirus Nei1 similarly lacks glycosylase activity. A structure of a G86D-DNA complex reveals a rearrangement in the β4/5 loop comprising Leu84, the highly-conserved void-filling residue, thereby providing a structural rationale for the decreased glycosylase activity of the glycine to aspartate variant.
KW - Base excision repair
KW - DNA glycosylase
KW - NEIL1
KW - Oxidized DNA lesions
KW - Single nucleotide polymorphisms
UR - http://www.scopus.com/inward/record.url?scp=84892824761&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84892824761&partnerID=8YFLogxK
U2 - 10.1016/j.dnarep.2013.12.003
DO - 10.1016/j.dnarep.2013.12.003
M3 - Article
C2 - 24382305
AN - SCOPUS:84892824761
SN - 1568-7864
VL - 14
SP - 17
EP - 26
JO - DNA Repair
JF - DNA Repair
IS - 1
ER -