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Journal of Virology, August 2003, p. 9052-9068, Vol. 77, No. 16
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.16.9052-9068.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Redistribution of Cyclophilin A to Viral Factories during Vaccinia Virus Infection and Its Incorporation into Mature Particles

Ana Paula V. Castro,1 Técia M. U. Carvalho,2 Nissin Moussatché,1 and Clarissa R. A. Damaso1*

Laboratório de Biologia Molecular de Vírus,1 Laboratório de Ultraestrutura Celular Hertha Meyer, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ 21941-590, Brazil2

Received 7 February 2003/ Accepted 28 May 2003

Cyclophilins are peptidyl-prolyl cis-trans isomerases involved in catalyzing conformational changes and accelerating the rate of protein folding and refolding in several cellular systems. In the present study, we analyzed the expression pattern and intracellular distribution of the cellular isomerase cyclophilin A (CypA) during vaccinia virus (VV) infection. An impressive increase in CypA stability was observed, leading to a practically unchanged accumulation of CypA during infection, although its synthesis was completely inhibited at late times. By confocal microscopy, we observed that CypA went through an intense reorganization in the cell cytoplasm and colocalized with the virosomes late in infection. CypA relocation to viral factories required the synthesis of viral postreplicative proteins, and treatment of infected cells with cyclosporine (CsA) prevented CypA relocation, clearly excluding the virosomes from CypA staining. Immunoelectron microscopy of VV-infected cells showed that CypA was incorporated into VV particles during morphogenesis. Biochemical and electron microscopic assays with purified virions confirmed that CypA was encapsidated within the virus particle and localized specifically in the core. This work suggests that CypA may develop an important role in VV replication.

* Corresponding author. Mailing address: Laboratorio de Biologia Molecular de Virus, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Av. Brigadeiro Trompovski s/n, CCS sala C1-028, Cidade Universitária, Rio de Janeiro, RJ 21941-590, Brazil. Phone: 55 21 2562-2433. Fax: 55 21 2280-8193. E-mail: damasoc{at}biof.ufrj.br.

Journal of Virology, August 2003, p. 9052-9068, Vol. 77, No. 16
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.16.9052-9068.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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