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Journal of Virology, January 2003, p. 97-104, Vol. 77, No. 1
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.1.97-104.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Heterodimerization of the Two Major Envelope Proteins Is Essential for Arterivirus Infectivity

Eric J. Snijder,* Jessika C. Dobbe, and Willy J. M. Spaan

Molecular Virology Laboratory, Department of Medical Microbiology, Center of Infectious Diseases, Leiden University Medical Center, Leiden, The Netherlands

Received 5 July 2002/ Accepted 24 September 2002

The two major envelope proteins of arteriviruses, the membrane protein (M) and the major glycoprotein (GP5), associate into a disulfide-linked heterodimer that is incorporated into the virion and has been assumed to be a prerequisite for virus assembly. Using an equine arteritis virus (EAV) infectious cDNA clone, we have analyzed the requirement for GP5-M heterodimerization and have identified the Cys residues involved in the formation of the GP5-M disulfide bond. The single Cys residue (Cys-8) in the M ectodomain was crucial for heterodimerization and virus infectivity. Mutagenesis of any of the five Cys residues in the GP5 ectodomain or removal of the single GP5 N-glycosylation site also rendered the full-length clone noninfectious. However, an analysis of revertants yielded an exceptional pseudorevertant in which residues 52 to 79 of the GP5 ectodomain had been deleted and the original Cys-80->Ser mutation had been maintained. Consequently, this revertant lacked the GP5 N-glycosyation site (Asn-56) and retained only a single cysteine residue (Cys-34). By using this GP5 deletion, we confirmed that Cys-34 of GP5 and Cys-8 of M are essential for GP5-M heterodimerization, a key event in the assembly of the EAV envelope.

* Corresponding author. Mailing address: Molecular Virology Laboratory, Department of Medical Microbiology, Leiden University Medical Center, LUMC E4-P, P.O. Box 9600, 2300 RC Leiden, The Netherlands. Phone: 31 71 5261657. Fax: 31 71 5266761. E-mail: E.J.Snijder{at}LUMC.nl.

Journal of Virology, January 2003, p. 97-104, Vol. 77, No. 1
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.1.97-104.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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