Miscoding events during DNA synthesis past the nitration-damaged base 8-nitroguanine

Naomi Suzuki, Manabu Yasui, Nicholas Geacintov, Vladimir Shafirovich, Shinya Shibutani

Research output: Contribution to journalArticle

Abstract

8-Nitro-2′-deoxyguanosine (8-NO2-dG) DNA adducts are induced by the reactive nitrogen species and may be associated with the development of cancer in inflammatory tissues. To explore the miscoding potential of 8-NO2-dG adduct, an oligodeoxynucleotide containing a single 8-NO2-dG adduct was prepared by photochemical synthesis and used as a template in primer extension reactions catalyzed by mammalian DNA polymerases (pol). Primer extension reactions catalyzed by pol α or β were strongly retarded at the 8-NO2-dG lesion; a fraction of primers was extended past the lesion by incorporating preferentially dCMP, the correct base, opposite the lesion, accompanied by lesser amounts of dAMP and dGMP incorporation. In contrast, primer extension reactions catalyzed by pol η or a truncated form of pol κ (pol κΔC) readily extended past the 8-NO2-dG lesion. Pol η and κΔC snowed more broad miscoding spectra; direct incorporations of dCMP and dAMP were observed, along with lesser amounts of dGMP and dTMP incorporations and deletions. The miscoding frequencies induced by pol η and κΔC were at least 8 times higher than that of pol α or β. Miscoding frequency and specificity of 8-NO2-dG varied depending on the DNA polymerases used. These observations were supported by steady-state kinetic studies. 8-NO2-dG adduct may play an important role in initiating inflammation driven carcinogenesis.

Original languageEnglish (US)
Pages (from-to)9238-9245
Number of pages8
JournalBiochemistry
Volume44
Issue number25
DOIs
StatePublished - Jun 28 2005

Fingerprint

Nitration
DNA
DNA-Directed DNA Polymerase
Reactive Nitrogen Species
DNA Adducts
Oligodeoxyribonucleotides
8-nitroguanine
8-nitro-2'-deoxyguanosine
Carcinogenesis
Tissue
Inflammation
Kinetics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Miscoding events during DNA synthesis past the nitration-damaged base 8-nitroguanine. / Suzuki, Naomi; Yasui, Manabu; Geacintov, Nicholas; Shafirovich, Vladimir; Shibutani, Shinya.

In: Biochemistry, Vol. 44, No. 25, 28.06.2005, p. 9238-9245.

Research output: Contribution to journalArticle

Suzuki, Naomi ; Yasui, Manabu ; Geacintov, Nicholas ; Shafirovich, Vladimir ; Shibutani, Shinya. / Miscoding events during DNA synthesis past the nitration-damaged base 8-nitroguanine. In: Biochemistry. 2005 ; Vol. 44, No. 25. pp. 9238-9245.
@article{57f291d599a7476cb767b50ad01b8146,
title = "Miscoding events during DNA synthesis past the nitration-damaged base 8-nitroguanine",
abstract = "8-Nitro-2′-deoxyguanosine (8-NO2-dG) DNA adducts are induced by the reactive nitrogen species and may be associated with the development of cancer in inflammatory tissues. To explore the miscoding potential of 8-NO2-dG adduct, an oligodeoxynucleotide containing a single 8-NO2-dG adduct was prepared by photochemical synthesis and used as a template in primer extension reactions catalyzed by mammalian DNA polymerases (pol). Primer extension reactions catalyzed by pol α or β were strongly retarded at the 8-NO2-dG lesion; a fraction of primers was extended past the lesion by incorporating preferentially dCMP, the correct base, opposite the lesion, accompanied by lesser amounts of dAMP and dGMP incorporation. In contrast, primer extension reactions catalyzed by pol η or a truncated form of pol κ (pol κΔC) readily extended past the 8-NO2-dG lesion. Pol η and κΔC snowed more broad miscoding spectra; direct incorporations of dCMP and dAMP were observed, along with lesser amounts of dGMP and dTMP incorporations and deletions. The miscoding frequencies induced by pol η and κΔC were at least 8 times higher than that of pol α or β. Miscoding frequency and specificity of 8-NO2-dG varied depending on the DNA polymerases used. These observations were supported by steady-state kinetic studies. 8-NO2-dG adduct may play an important role in initiating inflammation driven carcinogenesis.",
author = "Naomi Suzuki and Manabu Yasui and Nicholas Geacintov and Vladimir Shafirovich and Shinya Shibutani",
year = "2005",
month = "6",
day = "28",
doi = "10.1021/bi050276p",
language = "English (US)",
volume = "44",
pages = "9238--9245",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "25",

}

TY - JOUR

T1 - Miscoding events during DNA synthesis past the nitration-damaged base 8-nitroguanine

AU - Suzuki, Naomi

AU - Yasui, Manabu

AU - Geacintov, Nicholas

AU - Shafirovich, Vladimir

AU - Shibutani, Shinya

PY - 2005/6/28

Y1 - 2005/6/28

N2 - 8-Nitro-2′-deoxyguanosine (8-NO2-dG) DNA adducts are induced by the reactive nitrogen species and may be associated with the development of cancer in inflammatory tissues. To explore the miscoding potential of 8-NO2-dG adduct, an oligodeoxynucleotide containing a single 8-NO2-dG adduct was prepared by photochemical synthesis and used as a template in primer extension reactions catalyzed by mammalian DNA polymerases (pol). Primer extension reactions catalyzed by pol α or β were strongly retarded at the 8-NO2-dG lesion; a fraction of primers was extended past the lesion by incorporating preferentially dCMP, the correct base, opposite the lesion, accompanied by lesser amounts of dAMP and dGMP incorporation. In contrast, primer extension reactions catalyzed by pol η or a truncated form of pol κ (pol κΔC) readily extended past the 8-NO2-dG lesion. Pol η and κΔC snowed more broad miscoding spectra; direct incorporations of dCMP and dAMP were observed, along with lesser amounts of dGMP and dTMP incorporations and deletions. The miscoding frequencies induced by pol η and κΔC were at least 8 times higher than that of pol α or β. Miscoding frequency and specificity of 8-NO2-dG varied depending on the DNA polymerases used. These observations were supported by steady-state kinetic studies. 8-NO2-dG adduct may play an important role in initiating inflammation driven carcinogenesis.

AB - 8-Nitro-2′-deoxyguanosine (8-NO2-dG) DNA adducts are induced by the reactive nitrogen species and may be associated with the development of cancer in inflammatory tissues. To explore the miscoding potential of 8-NO2-dG adduct, an oligodeoxynucleotide containing a single 8-NO2-dG adduct was prepared by photochemical synthesis and used as a template in primer extension reactions catalyzed by mammalian DNA polymerases (pol). Primer extension reactions catalyzed by pol α or β were strongly retarded at the 8-NO2-dG lesion; a fraction of primers was extended past the lesion by incorporating preferentially dCMP, the correct base, opposite the lesion, accompanied by lesser amounts of dAMP and dGMP incorporation. In contrast, primer extension reactions catalyzed by pol η or a truncated form of pol κ (pol κΔC) readily extended past the 8-NO2-dG lesion. Pol η and κΔC snowed more broad miscoding spectra; direct incorporations of dCMP and dAMP were observed, along with lesser amounts of dGMP and dTMP incorporations and deletions. The miscoding frequencies induced by pol η and κΔC were at least 8 times higher than that of pol α or β. Miscoding frequency and specificity of 8-NO2-dG varied depending on the DNA polymerases used. These observations were supported by steady-state kinetic studies. 8-NO2-dG adduct may play an important role in initiating inflammation driven carcinogenesis.

UR - http://www.scopus.com/inward/record.url?scp=21744454850&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=21744454850&partnerID=8YFLogxK

U2 - 10.1021/bi050276p

DO - 10.1021/bi050276p

M3 - Article

VL - 44

SP - 9238

EP - 9245

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 25

ER -