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J Virol, April 1998, p. 2865-2870, Vol. 72, No. 4
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Role of Actin Microfilaments in Black Creek Canal Virus Morphogenesis

Eugene V. Ravkov,1 Stuart T. Nichol,2 Clarence J. Peters,2 and Richard W. Compans1,*

Department of Microbiology and Immunology, School of Medicine, Emory University, Atlanta, Georgia 30322,1 and Special Pathogens Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 303332

Received 18 September 1997/Accepted 17 December 1997

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 beta -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.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, School of Medicine, Emory University, 3001 Rollins Research Center, Atlanta, GA 30322. Phone: (404) 727-5947. Fax: (404) 727-8250.




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