title: Analysis of the interactions of viral and cellular factors with human cytomegalovirus lytic origin of replication, oriLyt.
abstract: Human cytomegalovirus transient lytic DNA replication relies on the cis-acting element oriLyt, six viral-encoded core proteins, the proposed DNA replication initiator protein UL84, IE2, IRS1 and the gene products from the UL112/113 loci. In an effort to elucidate cellular and viral-encoded factors that may play a role in oriLyt-dependent replication we used DNA-affinity purification and mass spectrometry to isolate and identify several previously unknown cellular and viral factors that interact with HCMV oriLyt DNA. These proteins include the multifunctional hnRNP-K, BUB3, HMGB1, PTB-1, UL83, UL112/113, and IRS1. Chromatin immunoprecipitation (ChIP) assays confirmed an interaction of several of these factors with oriLyt. Co-immunoprecipitation experiments detected an interaction between UL84 and hnRNP-K in infected and transfected cells. Knockdown of hnRNP K expression by siRNA inhibited the amplification of oriLyt in the transient assay. Together, these data suggest a possible regulatory role in DNA replication for several previously unidentified viral and cellular factors.
authors.affiliation: University of Nevada, Reno School of Medicine, Department of Microbiology & Immunology, Reno, NV 89557, USA.
authors.forename: Dominique
authors.initials: D
authors.lastname: Kagele
authors.forename: Cyprian C
authors.initials: CC
authors.lastname: Rossetto
authors.forename: Margaret T
authors.initials: MT
authors.lastname: Tarrant
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Pari
mesh_terms: D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics / Q000502:physiology*; D003586:Cytomegalovirus Infections / Q000378:metabolism* / Q000821:virology*; D004261:DNA Replication; D054884:Host-Pathogen Interactions; D006801:Humans; D000075223:Poly-ADP-Ribose Binding Proteins; D038941:Polypyrimidine Tract-Binding Protein / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D018741:Replication Origin; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: C489444:BUB3 protein, human; D018797:Cell Cycle Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D014764:Viral Proteins; C112091:oriLyt protein, Human cytomegalovirus; D038941:Polypyrimidine Tract-Binding Protein