title: The herpes simplex virus type 1 UL8 protein influences the intracellular localization of the UL52 but not the ICP8 or POL replication proteins in virus-infected cells.
abstract: We have developed a panel of 14 monoclonal antibodies (MAbs) to POL, the catalytic subunit of herpes simplex virus type 1 (HSV-1) DNA polymerase encoded by gene UL30, and one MAb to the UL52 protein, another of the seven proteins essential for replication of HSV DNA. The approximate locations of the epitopes of the polymerase-specific MAbs were identified using truncated polymerase molecules, and the antibodies were characterized in a number of immunological assays allowing eight different specificities to be recognized. These MAbs, together with a polyclonal antibody raised in rabbits against a third DNA replication protein, ICP8, were used to localize the respective proteins by immunofluorescence in cells infected with wild-type HSV-1 or the DNA replication-defective mutants ambUL8 or 2-2. In BHK cells infected with ambUL8, a mutant with an amber termination codon within the coding region of gene UL8, the UL52 protein did not enter the nucleus, although ICP8 and POL entered the nucleus in a normal fashion. The failure of the UL52 protein to be correctly transported to the nucleus was also observed in both HFL and Vero cells infected with ambUL8. In contrast, UL52 protein was transported to the nucleus in BHK cells infected with wild-type HSV-1 or with 2-2, a mutant lacking a functional UL9 protein.
authors.affiliation: MRC Virology Unit, Church Street, Glasgow, UK. H.Marsden@vir.gla.ac.uk
authors.forename: H S
authors.initials: HS
authors.lastname: Marsden
authors.forename: A M
authors.initials: AM
authors.lastname: Cross
authors.forename: G J
authors.initials: GJ
authors.lastname: Francis
authors.forename: A H
authors.initials: AH
authors.lastname: Patel
authors.forename: K
authors.initials: K
authors.lastname: MacEachran
authors.forename: M
authors.initials: M
authors.lastname: Murphy
authors.forename: G
authors.initials: G
authors.lastname: McVey
authors.forename: D
authors.initials: D
authors.lastname: Haydon
authors.forename: A
authors.initials: A
authors.lastname: Abbotts
authors.forename: N D
authors.initials: ND
authors.lastname: Stow
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D001692:Biological Transport; D002460:Cell Line; D002522:Chlorocebus aethiops; D006224:Cricetinae; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism*; D019915:DNA Primase; D004268:DNA-Binding Proteins; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D003673:Defective Viruses / Q000378:metabolism; D005090:Exodeoxyribonucleases; D017353:Gene Deletion; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D051379:Mice; D011817:Rabbits; D014709:Vero Cells; D014764:Viral Proteins / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D004268:DNA-Binding Proteins; C045462:ICP8 protein, Simplexvirus; D014764:Viral Proteins; D019915:DNA Primase; D004259:DNA-Directed DNA Polymerase; D005090:Exodeoxyribonucleases; C062021:helicase-primase, Human herpesvirus 1; C043417:DNA polymerase, Simplexvirus; D004265:DNA Helicases