title: Agnoprotein of polyomavirus BK interacts with proliferating cell nuclear antigen and inhibits DNA replication.
abstract: BACKGROUND
  The human polyomavirus BK expresses a 66 amino-acid peptide referred to as agnoprotein. Though mutants lacking agnoprotein are severely reduced in producing infectious virions, the exact function of this peptide remains incompletely understood. To elucidate the function of agnoprotein, we searched for novel cellular interaction partners.
  
  
  METHODS
  Yeast-two hybrid assay was performed with agnoprotein as bait against human kidney and thymus libraries. The interaction between agnoprotein and putative partners was further examined by GST pull down, co-immunoprecipitation, and fluorescence resonance energy transfer studies. Biochemical and biological studies were performed to examine the functional implication of the interaction of agnoprotein with cellular target proteins.
  
  
  RESULTS
  Proliferating cell nuclear antigen (PCNA), which acts as a processivity factor for DNA polymerase δ, was identified as an interaction partner. The interaction between agnoprotein and PCNA is direct and occurs also in human cells. Agnoprotein exerts an inhibitory effect on PCNA-dependent DNA synthesis in vitro and reduces cell proliferation when ectopically expressed. Overexpression of PCNA restores agnoprotein-mediated inhibition of cell proliferation.
  
  
  CONCLUSIONS
  Our data suggest that PCNA is a genuine interaction partner of agnoprotein and the inhibitory effect on PCNA-dependent DNA synthesis by the agnoprotein may play a role in switching off (viral) DNA replication late in the viral replication cycle when assembly of replicated genomes and synthesized viral capsid proteins occurs.
authors: Nancy
authors: N
authors: Gerits
authors: Mona
authors: M
authors: Johannessen
authors: Conny
authors: C
authors: Tümmler
authors: Mari
authors: M
authors: Walquist
authors: Sergiy
authors: S
authors: Kostenko
authors: Igor
authors: I
authors: Snapkov
authors: Barbara
authors: B
authors: van Loon
authors: Elena
authors: E
authors: Ferrari
authors: Ulrich
authors: U
authors: Hübscher
authors: UiT - The Arctic University of Norway, Faculty of Health Sciences, Department of Medical Biology, Molecular Inflammation Research Group, Tromsø NO-9037, Norway. ugo.moens@uit.no.
authors: Ugo
authors: U
authors: Moens
mesh_terms: D001739:BK Virus / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004258:DNA Polymerase III / Q000235:genetics / Q000378:metabolism; D004261:DNA Replication; D006801:Humans; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication; D000086082:Viroporin Proteins
chemical_list: D004258:DNA Polymerase III; D018809:Proliferating Cell Nuclear Antigen; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C042559:agnoprotein, polyomavirus; D000086082:Viroporin Proteins
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