Reconstitution of uracil DNA glcosylase-initiated base excision repair in herpes simplex virus-1

Federica Bogani, Chian New Chua, Paul E Boehmer

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Herpes simplex virus-1 is a large double-stranded DNA virus that is self-sufficient in a number of genome transactions. Hence, the virus encodes its own DNA replication apparatus and is capable of mediating recombination reactions. We recently reported that the catalytic subunit of the HSV-1 DNA polymerase (UL30) exhibits apurinic/apyrimidinic and 5′-deoxyribose phosphate lyase activities that are integral to base excision repair. Base excision repair is required to maintain genome stability as a means to counter the accumulation of unusual bases and to protect from the loss of DNA bases. Here we have reconstituted a system with purified HSV-1 and human proteins that perform all the steps of uracil DNA glycosylase-initiated base excision repair. In this system nucleotide incorporation is dependent on the HSV-1 uracil DNA glycosylase (UL2), human AP endonuclease, and the HSV-1 DNA polymerase. Completion of base excision repair can be mediated by T4 DNA ligase as well as human DNA ligase I or ligase IIIα-XRCC1 complex. Of these, ligase IIIα-XRCC1 is the most efficient. Moreover, ligase IIIα-XRCC1 confers specificity onto the reaction in as much as it allows ligation to occur in the presence of the HSV-1 DNA polymerase processivity factor (UL42) and prevents base excision repair from occurring with heterologous DNA polymerases. Completion of base excision repair in this system is also dependent on the incorporation of the correct nucleotide. These findings demonstrate that the HSV-1 proteins in combination with cellular factors that are not encoded by the virus are capable of performing base excision repair. These results have implications on the role of base excision repair in viral genome maintenance during lytic replication and reactivation from latency.

Original languageEnglish (US)
Pages (from-to)16784-16790
Number of pages7
JournalJournal of Biological Chemistry
Volume284
Issue number25
DOIs
StatePublished - Jun 19 2009

Fingerprint

Uracil
Human Herpesvirus 1
Viruses
DNA Repair
Repair
DNA
DNA-Directed DNA Polymerase
Ligases
Uracil-DNA Glycosidase
DNA Ligases
Genes
Nucleotides
DNA Viruses
Genomic Instability
Viral Genome
DNA Replication
Genetic Recombination
Ligation
Catalytic Domain
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Reconstitution of uracil DNA glcosylase-initiated base excision repair in herpes simplex virus-1. / Bogani, Federica; Chua, Chian New; Boehmer, Paul E.

In: Journal of Biological Chemistry, Vol. 284, No. 25, 19.06.2009, p. 16784-16790.

Research output: Contribution to journalArticle

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