Effect of manganese on in vitro replication of damaged DNA catalyzed by the herpes simplex virus type-1 DNA polymerase

Giuseppe Villani, Nicolas Tanguy Le Gac, Luc Wasungu, Dominique Burnouf, Robert P. Fuchs, Paul E Boehmer

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

In vitro bypass of damaged DNA by replicative DNA polymerases is usually blocked by helix-distorting or bulky DNA lesions. In this study, we report that substitution of the divalent metal ion Mg2+ with Mn2+ promotes quantitative replication of model DNA substrates containing the major cisplatin or N-2-acetylaminofluorene adducts by the catalytic subunit (UL30) of the replicative DNA polymerase of herpes simplex virus. The ability of Mn2+ ions to confer bypass of bulky lesions was not observed with other replicative DNA polymerases of the B family, such as bacteriophage T4 or δ polymerases. However, for these enzymes, manganese induced the incorporation of one nucleotide opposite the first (3′) guanine of the d(GpG) intrastrand cisplatin lesion. Translesion replication of the cisplatin adduct by UL30 led to the incorporation of mismatched bases, with the preferential incorporation of dAMP opposite the 3′ guanine of the lesion. Furthermore, substitution of MgCl2 with MnCl2 greatly inhibited the 3′ to 5′ exonuclease of UL30 but had a far lesser effect on that of T4 DNA polymerase. Finally, manganese induced a conformational change in the structure of UL30 bound to the platinated substrate. Taken together, the latter findings suggest a mechanism by which manganese might allow UL30 to efficiently promote translesion DNA synthesis in vitro.

Original languageEnglish (US)
Pages (from-to)3323-3332
Number of pages10
JournalNucleic Acids Research
Volume30
Issue number15
StatePublished - Aug 1 2002
Externally publishedYes

Fingerprint

Human Herpesvirus 1
DNA-Directed DNA Polymerase
Manganese
DNA Replication
Guanine
Cisplatin
DNA
Ions
2-Acetylaminofluorene
Bacteriophage T4
Exonucleases
Magnesium Chloride
Catalytic Domain
Nucleotides
Metals
Enzymes
Simplexvirus DNA polymerase
In Vitro Techniques

ASJC Scopus subject areas

  • Genetics

Cite this

Effect of manganese on in vitro replication of damaged DNA catalyzed by the herpes simplex virus type-1 DNA polymerase. / Villani, Giuseppe; Le Gac, Nicolas Tanguy; Wasungu, Luc; Burnouf, Dominique; Fuchs, Robert P.; Boehmer, Paul E.

In: Nucleic Acids Research, Vol. 30, No. 15, 01.08.2002, p. 3323-3332.

Research output: Contribution to journalArticle

Villani, Giuseppe ; Le Gac, Nicolas Tanguy ; Wasungu, Luc ; Burnouf, Dominique ; Fuchs, Robert P. ; Boehmer, Paul E. / Effect of manganese on in vitro replication of damaged DNA catalyzed by the herpes simplex virus type-1 DNA polymerase. In: Nucleic Acids Research. 2002 ; Vol. 30, No. 15. pp. 3323-3332.
@article{0fb15763ea6f4af3b9ad1ef5842d9644,
title = "Effect of manganese on in vitro replication of damaged DNA catalyzed by the herpes simplex virus type-1 DNA polymerase",
abstract = "In vitro bypass of damaged DNA by replicative DNA polymerases is usually blocked by helix-distorting or bulky DNA lesions. In this study, we report that substitution of the divalent metal ion Mg2+ with Mn2+ promotes quantitative replication of model DNA substrates containing the major cisplatin or N-2-acetylaminofluorene adducts by the catalytic subunit (UL30) of the replicative DNA polymerase of herpes simplex virus. The ability of Mn2+ ions to confer bypass of bulky lesions was not observed with other replicative DNA polymerases of the B family, such as bacteriophage T4 or δ polymerases. However, for these enzymes, manganese induced the incorporation of one nucleotide opposite the first (3′) guanine of the d(GpG) intrastrand cisplatin lesion. Translesion replication of the cisplatin adduct by UL30 led to the incorporation of mismatched bases, with the preferential incorporation of dAMP opposite the 3′ guanine of the lesion. Furthermore, substitution of MgCl2 with MnCl2 greatly inhibited the 3′ to 5′ exonuclease of UL30 but had a far lesser effect on that of T4 DNA polymerase. Finally, manganese induced a conformational change in the structure of UL30 bound to the platinated substrate. Taken together, the latter findings suggest a mechanism by which manganese might allow UL30 to efficiently promote translesion DNA synthesis in vitro.",
author = "Giuseppe Villani and {Le Gac}, {Nicolas Tanguy} and Luc Wasungu and Dominique Burnouf and Fuchs, {Robert P.} and Boehmer, {Paul E}",
year = "2002",
month = "8",
day = "1",
language = "English (US)",
volume = "30",
pages = "3323--3332",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "15",

}

TY - JOUR

T1 - Effect of manganese on in vitro replication of damaged DNA catalyzed by the herpes simplex virus type-1 DNA polymerase

AU - Villani, Giuseppe

AU - Le Gac, Nicolas Tanguy

AU - Wasungu, Luc

AU - Burnouf, Dominique

AU - Fuchs, Robert P.

AU - Boehmer, Paul E

PY - 2002/8/1

Y1 - 2002/8/1

N2 - In vitro bypass of damaged DNA by replicative DNA polymerases is usually blocked by helix-distorting or bulky DNA lesions. In this study, we report that substitution of the divalent metal ion Mg2+ with Mn2+ promotes quantitative replication of model DNA substrates containing the major cisplatin or N-2-acetylaminofluorene adducts by the catalytic subunit (UL30) of the replicative DNA polymerase of herpes simplex virus. The ability of Mn2+ ions to confer bypass of bulky lesions was not observed with other replicative DNA polymerases of the B family, such as bacteriophage T4 or δ polymerases. However, for these enzymes, manganese induced the incorporation of one nucleotide opposite the first (3′) guanine of the d(GpG) intrastrand cisplatin lesion. Translesion replication of the cisplatin adduct by UL30 led to the incorporation of mismatched bases, with the preferential incorporation of dAMP opposite the 3′ guanine of the lesion. Furthermore, substitution of MgCl2 with MnCl2 greatly inhibited the 3′ to 5′ exonuclease of UL30 but had a far lesser effect on that of T4 DNA polymerase. Finally, manganese induced a conformational change in the structure of UL30 bound to the platinated substrate. Taken together, the latter findings suggest a mechanism by which manganese might allow UL30 to efficiently promote translesion DNA synthesis in vitro.

AB - In vitro bypass of damaged DNA by replicative DNA polymerases is usually blocked by helix-distorting or bulky DNA lesions. In this study, we report that substitution of the divalent metal ion Mg2+ with Mn2+ promotes quantitative replication of model DNA substrates containing the major cisplatin or N-2-acetylaminofluorene adducts by the catalytic subunit (UL30) of the replicative DNA polymerase of herpes simplex virus. The ability of Mn2+ ions to confer bypass of bulky lesions was not observed with other replicative DNA polymerases of the B family, such as bacteriophage T4 or δ polymerases. However, for these enzymes, manganese induced the incorporation of one nucleotide opposite the first (3′) guanine of the d(GpG) intrastrand cisplatin lesion. Translesion replication of the cisplatin adduct by UL30 led to the incorporation of mismatched bases, with the preferential incorporation of dAMP opposite the 3′ guanine of the lesion. Furthermore, substitution of MgCl2 with MnCl2 greatly inhibited the 3′ to 5′ exonuclease of UL30 but had a far lesser effect on that of T4 DNA polymerase. Finally, manganese induced a conformational change in the structure of UL30 bound to the platinated substrate. Taken together, the latter findings suggest a mechanism by which manganese might allow UL30 to efficiently promote translesion DNA synthesis in vitro.

UR - http://www.scopus.com/inward/record.url?scp=0036682441&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0036682441&partnerID=8YFLogxK

M3 - Article

C2 - 12140316

AN - SCOPUS:0036682441

VL - 30

SP - 3323

EP - 3332

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 15

ER -