title: Association between the herpes simplex virus-1 DNA polymerase and uracil DNA glycosylase.
abstract: Herpes simplex virus-1 (HSV-1) is a large dsDNA virus that encodes its own DNA replication machinery and other enzymes involved in DNA transactions. We recently reported that the HSV-1 DNA polymerase catalytic subunit (UL30) exhibits apurinic/apyrimidinic and 5'-deoxyribose phosphate lyase activities. Moreover, UL30, in conjunction with the viral uracil DNA glycosylase (UL2), cellular apurinic/apyrimidinic endonuclease, and DNA ligase IIIalpha-XRCC1, performs uracil-initiated 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 show that the HSV-1 UL2 associates with the viral replisome. We identified UL2 as a protein that co-purifies with the DNA polymerase through numerous chromatographic steps, an interaction that was verified by co-immunoprecipitation and direct binding studies. The interaction between UL2 and the DNA polymerase is mediated through the UL30 subunit. Moreover, UL2 co-localizes with UL30 to nuclear viral prereplicative sites. The functional consequence of this interaction is that replication of uracil-containing templates stalls at positions -1 and -2 relative to the template uracil because of the fact that these are converted into non-instructional abasic sites. These findings support the existence of a viral repair complex that may be capable of replication-coupled base excision repair and further highlight the role of DNA repair in the maintenance of the HSV-1 genome.
authors.affiliation: Department of Basic Medical Sciences, The University of Arizona College of Medicine, Phoenix, Arizona 85004, USA.
authors.forename: Federica
authors.initials: F
authors.lastname: Bogani
authors.forename: Ilsa
authors.initials: I
authors.lastname: Corredeira
authors.forename: Virneliz
authors.initials: V
authors.lastname: Fernandez
authors.forename: Ulrike
authors.initials: U
authors.lastname: Sattler
authors.forename: Wiriya
authors.initials: W
authors.lastname: Rutvisuttinunt
authors.forename: Martine
authors.initials: M
authors.lastname: Defais
authors.forename: Paul E
authors.initials: PE
authors.lastname: Boehmer
mesh_terms: D020134:Catalytic Domain; D004260:DNA Repair; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D016679:Genome, Viral / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D009097:Multienzyme Complexes / Q000378:metabolism; D011485:Protein Binding; D021381:Protein Transport; D051981:Uracil-DNA Glycosidase / Q000302:isolation & purification / Q000378:metabolism*; D014779:Virus Replication; D000099272:Replisomes
chemical_list: D004279:DNA, Viral; D004259:DNA-Directed DNA Polymerase; D009097:Multienzyme Complexes; D051981:Uracil-DNA Glycosidase; D000099272:Replisomes