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J Virol, April 1998, p. 2655-2662, Vol. 72, No. 4
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Involvement of Actin Microfilaments in the Replication of Human Parainfluenza Virus Type 3

Sanhita Gupta,1 Bishnu P. De,2 Judith A. Drazba,3 and Amiya K. Banerjee1,2,*

Molecular Virology Graduate Program, Department of Biochemistry, Case Western Reserve University, Cleveland, Ohio 44106,1 and Departments of Molecular Biology2 and Neurosciences,3 The Lerner Research Institute, The Cleveland Clinic Foundation, Cleveland, Ohio 44195

Received 15 August 1997/Accepted 2 December 1997

Several studies indicate that paramyxoviruses require a specific cellular factor(s) for transcription of their genomic RNAs. We previously reported that the cellular cytoskeletal protein actin, in its polymeric form, participates in the transcription of human parainfluenza virus type 3 (HPIV3) in vitro. In the present study, we investigated the role of the polymeric form of actin, i.e., the actin microfilaments of the cytoskeletal framework, in the reproduction of HPIV3 in vivo. Pulse-chase labeling analyses indicate that the viral nucleocapsid-associated proteins, NP and P, are present predominantly in the cytoskeletal framework during infection. By in situ hybridization, we found that viral mRNAs and genomic RNA were synthesized from the nucleocapsids that were bound to the cytoskeletal framework. Double immunofluorescent labeling and confocal microscopy of the cytoarchitecture revealed that the viral nucleocapsids are specifically localized on the actin microfilaments. Treatment of cells with the actin-depolymerizing agent, cytochalasin D, resulted in the inhibition of viral RNA synthesis and ribonucleoprotein accumulation. These results strongly suggest that actin microfilaments play an important role in the replication of HPIV3.

* Corresponding author. Mailing address: Department of Molecular Biology, The Cleveland Clinic Foundation, 9500 Euclid Ave., NC20, Cleveland, OH 44195. Phone: (216) 444-0625. Fax: (216) 444-0512. E-mail: banerja{at}cesmtp.ccf.org.




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