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Journal of Virology, March 2000, p. 2949-2954, Vol. 74, No. 6
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Characterization of the R572T Point Mutant of a Putative Cleavage Site in Human Foamy Virus Env

Anju Bansal,1 Kit L. Shaw,2 Bradley H. Edwards,1 Paul A. Goepfert,1,2 and Mark J. Mulligan1,2,*

Departments of Medicine1 and Microbiology,2 University of Alabama at Birmingham, Birmingham, Alabama 35294-2170

Received 25 June 1999/Accepted 8 December 1999

A putative cleavage site of the human foamy virus (HFV) envelope glycoprotein (Env) was altered. Transient env expression revealed that the R572T mutant Env was normally expressed and modified by asparagine-linked oligosaccharide chains. However, this single-amino-acid substitution was sufficient to abolish all detectable cleavage of the gp130 precursor polyprotein. Cell surface biotinylation demonstrated that the uncleaved mutant gp130 was transported to the plasma membrane. The uncleaved mutant protein was incapable of syncytium formation. Glycoprotein-driven virion budding, a unique aspect of HFV assembly, occurred despite the absence of Env cleavage. We then substituted the R572T mutant env into a replication-competent HFV molecular clone. Transfection of the mutant viral DNA into BHK-21 cells followed by viral titration with the FAB (foamy virus-activated beta -galactosidase expression) assay revealed that proteolysis of the HFV Env was essential for viral infectivity. Wild-type HFV Env partially complemented the defective virus phenotype. Taken together, these experimental results established the location of the HFV Env proteolytic site; the effects of cleavage on Env transport, processing, and function; and the importance of Env proteolysis for virus maturation and infectivity.

* Corresponding author. Mailing address: Division of Infectious Diseases, 845 19th St. South, BBRB 220, University of Alabama at Birmingham, Birmingham, AL 35294-2170. Phone: (205) 975-8982. Fax: (205) 975-6027. E-mail: mulligan{at}uab.edu.

Journal of Virology, March 2000, p. 2949-2954, Vol. 74, No. 6
0022-538X/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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