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Regulator of G protein signalling 16 is a target for a porcine circovirus type 2 protein

¹ Department of Biomedical Sciences and Veterinary Public Health, Section of Immunology, Swedish University of Agricultural Sciences, Biomedical Center, Box 588, SE-751 23 Uppsala, Sweden
² Institute of Gene Technology, Tallinn University of Technology, Akadeemia tee 15, Tallinn 12618, Estonia
³ INRA, UMR1079, F-35000 Rennes, France
⁴ Department of Biomedical Sciences and Veterinary Public Health, Section of Parasitology and Virology, Swedish University of Agricultural Sciences, PO Box 7036, SE-750 07 Uppsala, Sweden

Correspondence
Sirje Timmusk
Sirje.Timmusk{at}bvf.slu.se

Interaction studies have suggested that the non-structural protein encoded by open reading frame 3 (ORF3) of porcine circovirus type 2 (PCV2) binds specifically to a regulator of G protein signalling (RGS) related to human RGS16 (huRGS16). The full-length clone of RGS16 was generated from porcine cells and sequence analysis revealed a close relationship to huRGS16 and murine RGS16. In vitro pull-down experiments verified an interaction between porcine RGS16 (poRGS16) and ORF3 from PCV2. Using GST-linked ORF3 proteins from three different genogroups of PCV2 and from porcine circovirus type 1 (PCV1) in the pull-down experiments indicated that there were differences in their ability to bind poRGS16. Quantitative RT-PCR demonstrated that the expression of poRGS16 mRNA could be induced by a number of cell activators including mitogens (LPS and PHA), interferon inducers (ODN 2216 and poly I : C) and the neurotransmitter norepinephrine. Immunofluorescence labelling confirmed the induced expression of poRGS16 at the protein level and suggested that the PCV2 ORF3 protein co-localized with poRGS16 in LPS-activated porcine PBMC. Furthermore, poRGS16 appeared to participate in the translocation of the ORF3 protein into the cell nucleus, suggesting that the observed interaction may play an important role in the infection biology of porcine circovirus.

The GenBank/EMBL/DDBJ accession number of the full-length clone of porcine RGS16 isolated from PK-15/A cells is EU271873.

Two supplementary figures, showing colocalization of porcine RGS16 and PCV2–ORF2 in PBMCs and PK15/A, and a supplementary table of primer sequences are available with the online version of this paper.