Agnoprotein of polyomavirus BK interacts with proliferating cell nuclear antigen and inhibits DNA replication

Virology Journal

Ugo Moens³  Ulrich Hübscher²  Elena Ferrari²  Barbara van Loon²  Igor Snapkov³  Sergiy Kostenko⁵  Mari Walquist¹  Conny Tümmler³  Mona Johannessen⁴  Nancy Gerits³ 

[1] Present address: UiT - The Arctic University of Norway, Faculty of Biosciences, Fisheries and Economics, Norwegian College of Fishery Sciences, Group of Seafood Science, Tromsø NO-9037, Norway;Institute of Veterinary Biochemistry and Molecular Biology, University of Zürich-Irchel, Winterthurerstrasse 190, Zürich 8057, Switzerland;UiT - The Arctic University of Norway, Faculty of Health Sciences, Department of Medical Biology, Molecular Inflammation Research Group, Tromsø NO-9037, Norway;Faculty of Health Sciences, Department of Medical Biology, Host Microbe Interaction Research Group, Tromsø NO-9037, Norway;Present address: University of Bergen, Department of Molecular Biology, Molecular Bioenergetics and Signalling, Bergen NO-5020, Norway

关键词: Polyomaviruses;    PCNA;    DNA replication;    DNA polymerase;    Cell proliferation;    Agnoprotein;   

Others  :  1131033
DOI  :  10.1186/s12985-014-0220-1

 received in 2014-02-06, accepted in 2014-12-01,  发布年份 2015

【 摘 要 】

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.

Conclusion

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.

【 授权许可】

   
2015 Gerits et al.; licensee BioMed Central.

【 图 表 】

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