Replication, gene expression and particle production by a consensus Merkel Cell Polyomavirus (MCPyV) genome.
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Replication, gene expression and particle production by a consensus Merkel Cell Polyomavirus (MCPyV) genome. / Neumann, Frederike; Borchert, Sophie; Schmidt, Claudia; Reimer, Rudolph; Hohenberg, Heinrich; Fischer, Nicole; Grundhoff, Adam.
in: PLOS ONE, Jahrgang 6, Nr. 12, 12, 2011, S. 29112.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Replication, gene expression and particle production by a consensus Merkel Cell Polyomavirus (MCPyV) genome.
AU - Neumann, Frederike
AU - Borchert, Sophie
AU - Schmidt, Claudia
AU - Reimer, Rudolph
AU - Hohenberg, Heinrich
AU - Fischer, Nicole
AU - Grundhoff, Adam
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
U2 - 10.1371/journal.pone.0029112
DO - 10.1371/journal.pone.0029112
M3 - SCORING: Journal article
VL - 6
SP - 29112
JO - PLOS ONE
JF - PLOS ONE
SN - 1932-6203
IS - 12
M1 - 12
ER -
