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Journal of Virology, October 1999, p. 8152-8159, Vol. 73, No. 10
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Amino Acid Substitutions within the Leucine Zipper
Domain of the Murine Coronavirus Spike Protein Cause Defects in
Oligomerization and the Ability To Induce Cell-to-Cell
Fusion
Zongli
Luo,
Avery M.
Matthews, and
Susan R.
Weiss*
Department of Microbiology, University of
Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6076
Received 1 March 1999/Accepted 6 July 1999
The murine coronavirus spike (S) protein contains a leucine zipper
domain which is highly conserved among coronaviruses. To assess the
role of this leucine zipper domain in S-induced cell-to-cell fusion,
the six heptadic leucine and isoleucine residues were replaced with
alanine by site-directed mutagenesis. The mutant S proteins were
analyzed for cell-to-cell membrane fusion activity as well as for
progress through the glycoprotein maturation process, including
intracellular glycosylation, oligomerization, and cell surface
expression. Single-alanine-substitution mutations had minimal, if any,
effects on S-induced cell-to-cell fusion. Significant reduction in
fusion activity was observed, however, when two of the four middle
heptadic leucine or isoleucine residues were replaced with alanine.
Double alanine substitutions that involved either of the two end
heptadic leucine residues did not significantly affect fusion. All
double-substitution mutant S proteins displayed levels of
endoglycosidase H resistance and cell surface expression similar to
those of the wild-type S. However, fusion-defective double-alanine-substitution mutants exhibited defects in S
oligomerization. These results indicate that the leucine zipper domain
plays a role in S-induced cell-to-cell fusion and that the ability of S
to induce fusion may be dependent on the oligomeric structure of S.
*
Corresponding author. Mailing address: Department of
Microbiology, University of Pennsylvania School of Medicine,
Philadelphia, PA 19104-6076. Phone: (215) 898-8013. Fax: (215)
573-4858. E-mail: weisssr{at}mail.med.upenn.edu.
Journal of Virology, October 1999, p. 8152-8159, Vol. 73, No. 10
0022-538X/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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