Human metapneumovirus Induces Reorganization of the Actin Cytoskeleton for Direct Cell-to-Cell Spread.
Paramyxovirus spread generally involves assembly of individual viral particles which then infect target cells. We show that infection of human bronchial airway cells with human metapneumovirus (HMPV), a recently identified paramyxovirus which causes significant respiratory disease, results in formation of intercellular extensions and extensive networks of branched cell-associated filaments. Formation of these structures is dependent on actin, but not microtubule, polymerization. Interestingly, using a co-culture assay we show that conditions which block regular infection by HMPV particles, including addition of neutralizing antibodies or removal of cell surface heparan sulfate, did not prevent viral spread from infected to new target cells. In contrast, inhibition of actin polymerization or alterations to Rho GTPase signaling pathways significantly decreased cell-to-cell spread. Furthermore, viral proteins and viral RNA were detected in intercellular extensions, suggesting direct transfer of viral genetic material to new target cells. While roles for paramyxovirus matrix and fusion proteins in membrane deformation have been previously demonstrated, we show that the HMPV phosphoprotein extensively co-localized with actin and induced formation of cellular extensions when transiently expressed, supporting a new model in which a paramyxovirus phosphoprotein is a key player in assembly and spread. Our results reveal a novel mechanism for HMPV direct cell-to-cell spread and provide insights into dissemination of respiratory viruses.
Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, United States of America.
Farah
F
El Najjar
Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, United States of America.
Nicolás
N
Cifuentes-Muñoz
Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, United States of America.
Jing
http://orcid.org/0000-0002-6238-6696
J
Chen
Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, United States of America.
Haining
H
Zhu
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland, United States of America.
Ursula J
UJ
Buchholz
Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, United States of America.
Carole L
CL
Moncman
Department of Molecular and Cellular Biochemistry, University of Kentucky, Lexington, Kentucky, United States of America.
Rebecca Ellis
RE
Dutch
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