Recognition of DNA adducts by edited and unedited forms of DNA glycosylase NEIL1

Irina G. Minko, Vladimir L. Vartanian, Naoto N. Tozaki, Erdem Coskun, Sanem Hosbas Coskun, Pawel Jaruga, Jongchan Yeo, Sheila S. David, Michael P. Stone, Martin Egli, Miral Dizdaroglu, Amanda K. McCullough, R. Stephen Lloyd

Research output: Contribution to journalArticle

Abstract

Pre-mRNA encoding human NEIL1 undergoes editing by adenosine deaminase ADAR1 that converts a single adenosine to inosine, and this conversion results in an amino acid change of lysine 242 to arginine. Previous investigations of the catalytic efficiencies of the two forms of the enzyme revealed differential release of thymine glycol (ThyGly) from synthetic oligodeoxynucleotides, with the unedited form, NEIL1 K242 being ≈30-fold more efficient than the edited NEIL1 K242R. In contrast, when these enzymes were reacted with oligodeoxynucleotides containing guanidinohydantoin or spiroiminohydantoin, the edited K242R form was ≈3-fold more efficient than the unedited NEIL1. However, no prior studies have investigated the efficiencies of these two forms of NEIL1 on either high-molecular weight DNA containing multiple oxidatively-induced base damages, or oligodeoxynucleotides containing a bulky alkylated formamidopyrimidine. To understand the extent of changes in substrate recognition, γ-irradiated calf thymus DNA was treated with either edited or unedited NEIL1 and the released DNA base lesions analyzed by gas chromatography-tandem mass spectrometry. Of all the measured DNA lesions, imidazole ring-opened 4,6-diamino-5-formamidopyrimidine (FapyAde) and 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua) were preferentially released by both NEIL1 enzymes with K242R being ≈1.3 and 1.2-fold more efficient than K242 on excision of FapyAde and FapyGua, respectively. Consistent with the prior literature, large differences (≈7.5 to 12-fold) were measured in the excision of ThyGly from genomic DNA by the unedited versus edited NEIL1. In contrast, the edited NEIL1 was more efficient (≈3 to 5-fold) on release of 5-hydroxycytosine. Excision kinetics on DNA containing a site-specific aflatoxin B1-FapyGua adduct revealed an ≈1.4-fold higher rate by the unedited NEIL1. Molecular modeling provides insight into these differential substrate specificities. The results of this study and in particular, the comparison of substrate specificities of unedited and edited NEIL1 using biologically and clinically important base lesions, are critical for defining its role in preservation of genomic integrity.

Original languageEnglish (US)
Article number102741
JournalDNA Repair
Volume85
DOIs
StatePublished - Jan 2020

Fingerprint

DNA Glycosylases
DNA Adducts
Oligodeoxyribonucleotides
DNA
Substrate Specificity
Substrates
Enzymes
Inosine
Aflatoxin B1
Adenosine Deaminase
Molecular modeling
RNA Precursors
Tandem Mass Spectrometry
Gas chromatography
Adenosine
Gas Chromatography-Mass Spectrometry
Lysine
Mass spectrometry
Arginine
Molecular Weight

Keywords

  • Aflatoxin
  • Base excision repair
  • Formamidopyrimidines
  • Hepatocellular carcinoma
  • RNA editing
  • Thymine glycol

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Minko, I. G., Vartanian, V. L., Tozaki, N. N., Coskun, E., Coskun, S. H., Jaruga, P., ... Lloyd, R. S. (2020). Recognition of DNA adducts by edited and unedited forms of DNA glycosylase NEIL1. DNA Repair, 85, [102741]. https://doi.org/10.1016/j.dnarep.2019.102741

Recognition of DNA adducts by edited and unedited forms of DNA glycosylase NEIL1. / Minko, Irina G.; Vartanian, Vladimir L.; Tozaki, Naoto N.; Coskun, Erdem; Coskun, Sanem Hosbas; Jaruga, Pawel; Yeo, Jongchan; David, Sheila S.; Stone, Michael P.; Egli, Martin; Dizdaroglu, Miral; McCullough, Amanda K.; Lloyd, R. Stephen.

In: DNA Repair, Vol. 85, 102741, 01.2020.

Research output: Contribution to journalArticle

Minko, IG, Vartanian, VL, Tozaki, NN, Coskun, E, Coskun, SH, Jaruga, P, Yeo, J, David, SS, Stone, MP, Egli, M, Dizdaroglu, M, McCullough, AK & Lloyd, RS 2020, 'Recognition of DNA adducts by edited and unedited forms of DNA glycosylase NEIL1', DNA Repair, vol. 85, 102741. https://doi.org/10.1016/j.dnarep.2019.102741
Minko IG, Vartanian VL, Tozaki NN, Coskun E, Coskun SH, Jaruga P et al. Recognition of DNA adducts by edited and unedited forms of DNA glycosylase NEIL1. DNA Repair. 2020 Jan;85. 102741. https://doi.org/10.1016/j.dnarep.2019.102741
Minko, Irina G. ; Vartanian, Vladimir L. ; Tozaki, Naoto N. ; Coskun, Erdem ; Coskun, Sanem Hosbas ; Jaruga, Pawel ; Yeo, Jongchan ; David, Sheila S. ; Stone, Michael P. ; Egli, Martin ; Dizdaroglu, Miral ; McCullough, Amanda K. ; Lloyd, R. Stephen. / Recognition of DNA adducts by edited and unedited forms of DNA glycosylase NEIL1. In: DNA Repair. 2020 ; Vol. 85.
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abstract = "Pre-mRNA encoding human NEIL1 undergoes editing by adenosine deaminase ADAR1 that converts a single adenosine to inosine, and this conversion results in an amino acid change of lysine 242 to arginine. Previous investigations of the catalytic efficiencies of the two forms of the enzyme revealed differential release of thymine glycol (ThyGly) from synthetic oligodeoxynucleotides, with the unedited form, NEIL1 K242 being ≈30-fold more efficient than the edited NEIL1 K242R. In contrast, when these enzymes were reacted with oligodeoxynucleotides containing guanidinohydantoin or spiroiminohydantoin, the edited K242R form was ≈3-fold more efficient than the unedited NEIL1. However, no prior studies have investigated the efficiencies of these two forms of NEIL1 on either high-molecular weight DNA containing multiple oxidatively-induced base damages, or oligodeoxynucleotides containing a bulky alkylated formamidopyrimidine. To understand the extent of changes in substrate recognition, γ-irradiated calf thymus DNA was treated with either edited or unedited NEIL1 and the released DNA base lesions analyzed by gas chromatography-tandem mass spectrometry. Of all the measured DNA lesions, imidazole ring-opened 4,6-diamino-5-formamidopyrimidine (FapyAde) and 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua) were preferentially released by both NEIL1 enzymes with K242R being ≈1.3 and 1.2-fold more efficient than K242 on excision of FapyAde and FapyGua, respectively. Consistent with the prior literature, large differences (≈7.5 to 12-fold) were measured in the excision of ThyGly from genomic DNA by the unedited versus edited NEIL1. In contrast, the edited NEIL1 was more efficient (≈3 to 5-fold) on release of 5-hydroxycytosine. Excision kinetics on DNA containing a site-specific aflatoxin B1-FapyGua adduct revealed an ≈1.4-fold higher rate by the unedited NEIL1. Molecular modeling provides insight into these differential substrate specificities. The results of this study and in particular, the comparison of substrate specificities of unedited and edited NEIL1 using biologically and clinically important base lesions, are critical for defining its role in preservation of genomic integrity.",
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author = "Minko, {Irina G.} and Vartanian, {Vladimir L.} and Tozaki, {Naoto N.} and Erdem Coskun and Coskun, {Sanem Hosbas} and Pawel Jaruga and Jongchan Yeo and David, {Sheila S.} and Stone, {Michael P.} and Martin Egli and Miral Dizdaroglu and McCullough, {Amanda K.} and Lloyd, {R. Stephen}",
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AU - Vartanian, Vladimir L.

AU - Tozaki, Naoto N.

AU - Coskun, Erdem

AU - Coskun, Sanem Hosbas

AU - Jaruga, Pawel

AU - Yeo, Jongchan

AU - David, Sheila S.

AU - Stone, Michael P.

AU - Egli, Martin

AU - Dizdaroglu, Miral

AU - McCullough, Amanda K.

AU - Lloyd, R. Stephen

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N2 - Pre-mRNA encoding human NEIL1 undergoes editing by adenosine deaminase ADAR1 that converts a single adenosine to inosine, and this conversion results in an amino acid change of lysine 242 to arginine. Previous investigations of the catalytic efficiencies of the two forms of the enzyme revealed differential release of thymine glycol (ThyGly) from synthetic oligodeoxynucleotides, with the unedited form, NEIL1 K242 being ≈30-fold more efficient than the edited NEIL1 K242R. In contrast, when these enzymes were reacted with oligodeoxynucleotides containing guanidinohydantoin or spiroiminohydantoin, the edited K242R form was ≈3-fold more efficient than the unedited NEIL1. However, no prior studies have investigated the efficiencies of these two forms of NEIL1 on either high-molecular weight DNA containing multiple oxidatively-induced base damages, or oligodeoxynucleotides containing a bulky alkylated formamidopyrimidine. To understand the extent of changes in substrate recognition, γ-irradiated calf thymus DNA was treated with either edited or unedited NEIL1 and the released DNA base lesions analyzed by gas chromatography-tandem mass spectrometry. Of all the measured DNA lesions, imidazole ring-opened 4,6-diamino-5-formamidopyrimidine (FapyAde) and 2,6-diamino-4-hydroxy-5-formamidopyrimidine (FapyGua) were preferentially released by both NEIL1 enzymes with K242R being ≈1.3 and 1.2-fold more efficient than K242 on excision of FapyAde and FapyGua, respectively. Consistent with the prior literature, large differences (≈7.5 to 12-fold) were measured in the excision of ThyGly from genomic DNA by the unedited versus edited NEIL1. In contrast, the edited NEIL1 was more efficient (≈3 to 5-fold) on release of 5-hydroxycytosine. Excision kinetics on DNA containing a site-specific aflatoxin B1-FapyGua adduct revealed an ≈1.4-fold higher rate by the unedited NEIL1. Molecular modeling provides insight into these differential substrate specificities. The results of this study and in particular, the comparison of substrate specificities of unedited and edited NEIL1 using biologically and clinically important base lesions, are critical for defining its role in preservation of genomic integrity.

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