Replication, gene expression and particle production by a consensus Merkel Cell Polyomavirus (MCPyV) genome

• Published in: PloS one Vol. 6; no. 12; p. e29112
• Main Authors: Neumann, Friederike, Borchert, Sophie, Schmidt, Claudia, Reimer, Rudolph, Hohenberg, Heinrich, Fischer, Nicole, Grundhoff, Adam
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 27.12.2011; Public Library of Science (PLoS)
• Subjects: Antigen T (large); Antigens; Biology; Biotechnology; Cancer; Cell cycle; Cell Line, Tumor; Cells (Biology); Cellulose; Cloning; Conserved sequence; Deoxyribonucleic acid; Deregulation; Dissection; DNA; DNA Replication; Gene Expression; Genes; Genome, Viral; Genomes; Genomics; Geriatrics; Humans; Infections; Life cycle analysis; Medicine; Merkel cell polyomavirus - genetics; Merkel cell polyomavirus - physiology; Mutation; Neuroblastoma; Older people; Particle formation; Particle production; Pathogenesis; Plasmids; Proteins; Replication; Skin; Tissues; Transcription; Transcription, Genetic; Transfection; Tumors; Virion; Virology; Virus Replication; Viruses; Germany
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0029112

Merkel Cell Polyomavirus (MCPyV) genomes are clonally integrated in tumor tissues of approximately 85% of all Merkel cell carcinoma (MCC) cases, a highly aggressive tumor of the skin which predominantly afflicts elderly and immunosuppressed patients. All integrated viral genomes recovered from MCC tissue or MCC cell lines harbor signature mutations in the early gene transcript encoding for the large T-Antigen (LT-Ag). These mutations selectively abrogate the ability of LT-Ag to support viral replication while still maintaining its Rb-binding activity, suggesting a continuous requirement for LT-Ag mediated cell cycle deregulation during MCC pathogenesis. To gain a better understanding of MCPyV biology, in vitro MCPyV replication systems are required. We have generated a synthetic MCPyV genomic clone (MCVSyn) based on the consensus sequence of MCC-derived sequences deposited in the NCBI database. Here, we demonstrate that transfection of recircularized MCVSyn DNA into some human cell lines recapitulates efficient replication of the viral genome, early and late gene expression together with virus particle formation. However, serial transmission of infectious virus was not observed. This in vitro culturing system allows the study of viral replication and will facilitate the molecular dissection of important aspects of the MCPyV lifecycle.