Abstract
BackgroundEleven new human polyomaviruses (HPyVs) have been identified in the last decade. Serological studies show that these novel HPyVs sub-clinically infect humans at an early age. The routes of infection, entry pathways, and cell tropism of new HPyVs remain unknown. VP1 proteins of polyomaviruses can assembly into virus-like particles (VLPs). As cell culturing systems for HPyV are currently not available, VP1-derived VLPs may be useful tools in basic research and biotechnological applications.ResultsRecombinant VP1-derived VLPs from 11 newly identified HPyVs were efficiently expressed in yeast. VP1 proteins derived from Merkel cell polyomavirus (MCPyV), trichodysplasia spinulosa-associated polyomavirus (TSPyV), and New Jersey polyomavirus (NJPyV) self-assembled into homogeneous similarly-sized VLPs. Karolinska Institutet polyomavirus (KIPyV), HPyV7, HPyV9, HPyV10, and St. Louis polyomavirus (STLPyV) VP1 proteins formed VLPs that varied in size with diameters ranging from 20 to 60 nm. Smaller-sized VLPs (25–35 nm in diameter) predominated in preparations from Washington University polyomavirus (WUPyV) and HPyV6. Attempts to express recombinant HPyV12 VP1-derived VLPs in yeast indicate that translation of VP1 might start at the second of two potential translation initiation sites in the VP1-encoding open reading frame (ORF). This translation resulted in a 364-amino acid-long VP1 protein, which efficiently self-assembled into typical PyV VLPs. MCPyV-, KIPyV-, TSPyV-, HPyV9-, HPyV10-, and HPyV12-derived VLPs showed hemagglutination (HA) assay activity in guinea pig erythrocytes, whereas WUPyV-, HPyV6-, HPyV7-, STLPyV- and NJPyV-derived VP1 VLPs did not.ConclusionsThe yeast expression system was successfully utilized for high-throughput production of recombinant VP1-derived VLPs from 11 newly identified HPyVs. HPyV12 VP1-derived VLPs were generated from the second of two potential translation initiation sites in the VP1-encoding ORF. Recombinant VLPs produced in yeast originated from different HPyVs demonstrated distinct HA activities and may be useful in virus diagnostics, capsid structure studies, or investigation of entry pathways and cell tropism of HPyVs until cell culture systems for new HPyVs are developed.
Highlights
Eleven new human polyomaviruses (HPyVs) have been identified in the last decade
We have previously demonstrated that Simian virus 40 (SV40), hamster polyomavirus (HaPyV), BK polyomavirus (BKPyV), JC polyomavirus (JCPyV), and murine polyomavirus (MPyV)-derived VP1 virus-like particles (VLPs) can be efficiently produced in the yeast (Saccharomyces cerevisiae) strain, AH22-214, using the pFX7 expression plasmid [29, 30]
We showed that the applied yeast system is suitable for efficient expression and selfassembly of VP1-derived VLPs from 11 novel HPyVs: Merkel cell polyomavirus (MCPyV), Karolinska Institutet polyomavirus (KIPyV), Washington University polyomavirus (WUPyV), HPyV6, HPyV7, trichodysplasia spinulosa-associated polyomavirus (TSPyV), HPyV9, Malawi polyomavirus (MWPyV), St. Louis polyomavirus (STLPyV), HPyV12, and New Jersey polyomavirus (NJPyV)
Summary
Eleven new human polyomaviruses (HPyVs) have been identified in the last decade. As a result of improved molecular diagnostic techniques, combined with strategies in sample preparation to enrich viral DNA content through reduction of host genomic DNA, 11 new HPyVs have been identified in the last decade. Of these novel HPyVs, Karolinska Institutet polyomavirus (KIPyV), Washington University polyomavirus (WUPyV), Norkiene et al BMC Biotechnology (2015) 15:68 and Merkel cell polyomavirus (MCPyV) were among the first to be discovered [7,8,9]. The latest virus, New Jersey polyomavirus (NJPyV), was discovered in endothelial cells, sites of myositis, and sites of cutaneous necrosis in a pancreatic transplant recipient [18]
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