Role of Actin Microfilaments in Black Creek Canal Virus Morphogenesis

ABSTRACT We have investigated the involvement of cytoskeletal proteins in the morphogenesis of Black Creek Canal virus (BCCV), a New World hantavirus. Immunofluorescent staining of BCCV-infected cells revealed a filamentous pattern of virus antigen, the appearance of which was sensitive to treatment with cytochalasin D, an actin microfilament-depolymerizing drug. Double immunofluorescence staining of BCCV-infected Vero cells with anti-BCCV nucleocapsid (N) monoclonal antibody and phalloidin revealed a colocalization of the BCCV N protein with actin microfilaments. A similar, though less prominent, filamentous pattern was observed in BHK21 cells transiently expressing the BCCV N protein alone but not in cells expressing the BCCV G1 and G2 glycoproteins. Moreover, the association of the N protein with actin microfilaments was confirmed by coimmunoprecipitation with β-actin-specific antibody. Treatment of the BCCV-infected Vero cells at 3 days postinfection with cytochalasin D decreased the yield of released BCCV by 94% relative to the yield from untreated cells. Pretreatment of Vero cells with cytochalasin D prior to and during BCCV adsorption and entry had no effect on the outcome of virus production. These results indicate that actin filaments may play an important role in hantavirus assembly and/or release.

[1]  D. Nayak,et al.  influenza virus-infected cells . proteins with cellular cytoskeletal elements in Association of influenza virus NP and M 1 , 1996 .

[2]  R. Compans,et al.  Polarized entry and release in epithelial cells of Black Creek Canal virus, a New World hantavirus , 1997, Journal of virology.

[3]  D. Phillips,et al.  Directional budding of human immunodeficiency virus from monocytes , 1996, Journal of virology.

[4]  R. Pettersson,et al.  Synthesis, Assembly, and Intracellular Transport of Bunyaviridae Membrane Proteins , 1996 .

[5]  C. Schmaljohn.  Molecular Biology of Hantaviruses , 1996 .

[6]  R. Greenman,et al.  Hantavirus pulmonary syndrome in Florida: association with the newly identified Black Creek Canal virus. , 1996, The American journal of medicine.

[7]  M Heinlein,et al.  Interaction of Tobamovirus Movement Proteins with the Plant Cytoskeleton , 1995, Science.

[8]  P. Cossart,et al.  Actin-based motility of vaccinia virus , 1995, Nature.

[9]  P. Rollin,et al.  Genetic and serologic analysis of Black Creek Canal virus and its association with human disease and Sigmodon hispidus infection. , 1995, Virology.

[10]  C. Sanderson,et al.  Interaction of Sendai viral F, HN, and M proteins with host cytoskeletal and lipid components in Sendai virus-infected BHK cells. , 1995, Virology.

[11]  G. Glass,et al.  Isolation of black creek canal virus, a new hantavirus from Sigmodon hispidus in Florida , 1995, Journal of medical virology.

[12]  K Foucar,et al.  Hantavirus pulmonary syndrome. Pathogenesis of an emerging infectious disease. , 1995, The American journal of pathology.

[13]  K. Simons,et al.  Involvement of microtubule motors in basolateral and apical transport in kidney cells , 1994, Nature.

[14]  D. Phillips,et al.  Role of the cytoskeleton in cell-to-cell transmission of human immunodeficiency virus , 1994, Journal of virology.

[15]  J. Childs,et al.  Genetic identification of a hantavirus associated with an outbreak of acute respiratory illness. , 1993, Science.

[16]  C. Rice,et al.  Sindbis virus expression vectors: packaging of RNA replicons by using defective helper RNAs , 1993, Journal of virology.

[17]  H. Klenk,et al.  Possible involvement of microtubule disruption in bipolar budding of a Sendai virus mutant, F1-R, in epithelial MDCK cells , 1993, Journal of virology.

[18]  S. Chen,et al.  Bunyavirus protein transport and assembly. , 1991, Current topics in microbiology and immunology.

[19]  R. Pettersson,et al.  Protein localization and virus assembly at intracellular membranes. , 1991, Current topics in microbiology and immunology.

[20]  H. Klenk,et al.  Altered budding site of a pantropic mutant of Sendai virus, F1-R, in polarized epithelial cells , 1990, Journal of virology.

[21]  S. Baker,et al.  Host cell proteins required for measles virus reproduction. , 1990, The Journal of general virology.

[22]  J. Smith,et al.  Immunoelectron microscopy of Rift Valley fever viral morphogenesis in primary rat hepatocytes. , 1987, Virology.

[23]  David E. Misek,et al.  Microtubules and actin filaments are not critically involved in the biogenesis of epithelial cell surface polarity , 1986, The Journal of cell biology.

[24]  G. Rutter,et al.  Involvement of actin filaments in budding of measles virus: studies on cytoskeletons of infected cells. , 1986, Virology.

[25]  R. Compans,et al.  Effects of hexose starvation and the role of sialic acid in influenza virus release. , 1983, Virology.

[26]  R. Compans,et al.  Effect of cytochalasin B on the maturation of enveloped viruses , 1979, The Journal of experimental medicine.

[27]  B. Sefton,et al.  The entry into host cells of Sindbis virus, vesicular stomatitis virus and Sendai virus , 1978, Cell.

[28]  U. K. Laemmli,et al.  Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 , 1970, Nature.

[29]  J. Duchin,et al.  Hantavirus Pulmonary Syndrome: A Clinical Description of 17 Patients with a Newly Recognized Disease , 1994 .