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Visualization of Borna Disease Virus Protein Interactions with Host Proteins using in situ Proximity Ligation Assay

Visualization of Borna Disease Virus Protein Interactions with Host Proteins using in situ Proximity Ligation Assay

Jonas Johansson Wensman1*a, Karl-Johan Leuchowius2,3 a, Jiting Yan1, Anna-Lena Berg4, Liv Bode5, Hanns Ludwig5, Sandor Belak6, Ulf Landegren2, Ola Soderberg2, Mikael Berg6

1Department of Clinical Sciences, Swedish University of Agricultural Sciences, P.O. Box 7054, SE-75007 Uppsala, Sweden; 2Department of Immunology, Genetics and Pathology, SciLifeLab, Uppsala University, Uppsala Biomedical Center, P.O. Box 815, SE-751 08 Uppsala, Sweden; 3Walter and Eliza Hall Institute of Medical Research, 1G Royal Parade, 3052 Parkville, Australia; 4Medical Products Agency, P.O. Box 26, SE-751 03 Uppsala, Sweden; 5Freelance Bornavirus Workgroup, Joint Senior Scientists, Beerenstr. 41, DE-14163 Berlin, Germany; 6Department of Biomedical Sciences and Veterinary Public Health, Swedish University of Agricultural Sciences, P.O. Box 7028, SE-750 07 Uppsala, Sweden.

ABSTRACT

Borna disease virus type 1 (BDV) comprises highly conserved neurotropic non-segmented negative strand RNA-virus variants causing neurological and behavioral disorders in a wide range of mammalian animals, possibly including humans. Viral persistence in the brain has been frequently observed, however, the exact mechanisms behind BDV’s ability to establish persistence despite a prominent immune response are not known. Here we have used in situ proximity ligation assay (in situ PLA), a selective tool for studying virus-host protein-protein interactions. BDV P (phosphoprotein) and N (nucleoprotein) have previously been reported to interact with several host proteins, thereby interfering with various signaling pathways. In this study, we focused on some of these interactions (BDV P-HMGB1, BDV N/P-Cdc2). First, we used rat glioma cell cultures persistently infected with a laboratory strain of BDV (C6BV ) to establish the assay. Next, in situ PLA was applied to detect BDV P in brain tissues of infected animals. Finally, protein-protein interactions were visualized in both C6BV and brain tissues of experimentally as well as naturally infected animals (rat and horse, respectively). BDV proteins and their interactions with host proteins could be shown in cell cultures (HMGB1, Cdc2) and in brain tissues of rat (HMGB1, Cdc2) and horse (Cdc2 only) infected with BDV. In this study, we have for the rst time directly visualized protein-protein interactions between BDV and its host, and thereby con rmed previous data to demonstrate ndings in cell cultures to be applicable also in experimentally and naturally infected animals.

 

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Hosts and Viruses

June

Vol.6, Iss.3, Pages 42-84

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