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Journal of Virology, November 2002, p. 11113-11122, Vol. 76, No. 21
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.21.11113-11122.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Differential Requirements for COPI Coats in Formation of Replication Complexes among Three Genera of Picornaviridae

Elena V. Gazina,1,2* Jason M. Mackenzie,3 Rebecca J. Gorrell,1,{dagger} and David A. Anderson1

Macfarlane Burnet Institute for Medical Research and Public Health, Melbourne, Victoria 3004,1 Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Herston, Queensland 4029, Australia,3 D. I. Ivanovsky Institute of Virology, Moscow 123098, Russia2

Received 19 February 2002/ Accepted 22 July 2002

Picornavirus RNA replication requires the formation of replication complexes (RCs) consisting of virus-induced vesicles associated with viral nonstructural proteins and RNA. Brefeldin A (BFA) has been shown to strongly inhibit RNA replication of poliovirus but not of encephalomyocarditis virus (EMCV). Here, we demonstrate that the replication of parechovirus 1 (ParV1) is partly resistant to BFA, whereas echovirus 11 (EV11) replication is strongly inhibited. Since BFA inhibits COPI-dependent steps in endoplasmic reticulum (ER)-Golgi transport, we tested a hypothesis that different picornaviruses may have differential requirements for COPI in the formation of their RCs. Using immunofluorescence and cryo-immunoelectron microscopy we examined the association of a COPI component, ß-COP, with the RCs of EMCV, ParV1, and EV11. EMCV RCs did not contain ß-COP. In contrast, ß-COP appeared to be specifically distributed to the RCs of EV11. In ParV1-infected cells ß-COP was largely dispersed throughout the cytoplasm, with some being present in the RCs. These results suggest that there are differences in the involvement of COPI in the formation of the RCs of various picornaviruses, corresponding to their differential sensitivity to BFA. EMCV RCs are likely to be formed immediately after vesicle budding from the ER, prior to COPI association with membranes. ParV1 RCs are formed from COPI-containing membranes but COPI is unlikely to be directly involved in their formation, whereas formation of EV11 RCs appears to be dependent on COPI association with membranes.

* Corresponding author. Mailing address: Macfarlane Burnet Institute for Medical Research and Public Health, GPO Box 2284, Melbourne, Victoria 3001, Australia. Phone: 61 3 9282 2236. Fax: 61 3 9282 2100. E-mail: gazina{at}burnet.edu.au.

{dagger} Present address: Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, Victoria 3052, Australia.

Journal of Virology, November 2002, p. 11113-11122, Vol. 76, No. 21
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.21.11113-11122.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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