title: Identification of rep-associated factors in herpes simplex virus type 1-induced adeno-associated virus type 2 replication compartments.
abstract: Adeno-associated virus (AAV) is a human parvovirus that replicates only in cells coinfected with a helper virus, such as adenovirus or herpes simplex virus type 1 (HSV-1). We previously showed that nine HSV-1 factors are able to support AAV rep gene expression and genome replication. To elucidate the strategy of AAV replication in the presence of HSV-1, we undertook a proteomic analysis of cellular and HSV-1 factors associated with Rep proteins and thus potentially recruited within AAV replication compartments (AAV RCs). This study resulted in the identification of approximately 60 cellular proteins, among which factors involved in DNA and RNA metabolism represented the largest functional categories. Validation analyses indicated that the cellular DNA replication enzymes RPA, RFC, and PCNA were recruited within HSV-1-induced AAV RCs. Polymerase delta was not identified but subsequently was shown to colocalize with Rep within AAV RCs even in the presence of the HSV-1 polymerase complex. In addition, we found that AAV replication is associated with the recruitment of components of the Mre11/Rad50/Nbs1 complex, Ku70 and -86, and the mismatch repair proteins MSH2, -3, and -6. Finally, several HSV-1 factors were also found to be associated with Rep, including UL12. We demonstrated for the first time that this protein plays a role during AAV replication by enhancing the resolution of AAV replicative forms and AAV particle production. Altogether, these analyses provide the basis to understand how AAV adapts its replication strategy to the nuclear environment induced by the helper virus.
authors.affiliation: INSERM U758, Lyon F-69007, France.
authors.forename: Armel
authors.initials: A
authors.lastname: Nicolas
authors.forename: Nathalie
authors.initials: N
authors.lastname: Alazard-Dany
authors.forename: Coline
authors.initials: C
authors.lastname: Biollay
authors.forename: Loredana
authors.initials: L
authors.lastname: Arata
authors.forename: Nelly
authors.initials: N
authors.lastname: Jolinon
authors.forename: Lauriane
authors.initials: L
authors.lastname: Kuhn
authors.forename: Myriam
authors.initials: M
authors.lastname: Ferro
authors.forename: Sandra K
authors.initials: SK
authors.lastname: Weller
authors.forename: Alberto L
authors.initials: AL
authors.lastname: Epstein
authors.forename: Anna
authors.initials: A
authors.lastname: Salvetti
authors.forename: Anna
authors.initials: A
authors.lastname: Greco
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics / Q000502:physiology*; D006367:HeLa Cells; D006561:Herpes Simplex / Q000378:metabolism* / Q000821:virology; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D010322:Parvoviridae Infections / Q000378:metabolism* / Q000821:virology; D011485:Protein Binding; D025941:Protein Interaction Mapping; D051716:Replication Protein A / Q000235:genetics / Q000378:metabolism; D051818:Replication Protein C / Q000235:genetics / Q000378:metabolism; D014709:Vero Cells; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D004268:DNA-Binding Proteins; C081632:RFC1 protein, human; C497167:RPA1 protein, human; D051716:Replication Protein A; D014764:Viral Proteins; C068734:rep proteins, Adeno-associated virus 2; D051818:Replication Protein C