N-glycans of F protein differentially affect fusion activity of human respiratory syncytial virus

Journal of Virology
Gert ZimmerGeorg Herrler

Abstract

The human respiratory syncytial virus (Long strain) fusion protein contains six potential N-glycosylation sites: N27, N70, N116, N120, N126, and N500. Site-directed mutagenesis of these positions revealed that the mature fusion protein contains three N-linked oligosaccharides, attached to N27, N70, and N500. By introducing these mutations into the F gene in different combinations, four more mutants were generated. All mutants, including a triple mutant devoid of any N-linked oligosaccharide, were efficiently transported to the plasma membrane, as determined by flow cytometry and cell surface biotinylation. None of the glycosylation mutations interfered with proteolytic activation of the fusion protein. Despite similar levels of cell surface expression, the glycosylation mutants affected fusion activity in different ways. While the N27Q mutation did not have an effect on syncytium formation, loss of the N70-glycan caused a fusion activity increase of 40%. Elimination of both N-glycans (N27/70Q mutant) reduced the fusion activity by about 50%. A more pronounced reduction of the fusion activity of about 90% was observed with the mutants N500Q, N27/500Q, and N70/500Q. Almost no fusion activity was detected with the triple mutant N2...Continue Reading

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Citations

Oct 15, 2003·Immunology Letters·Koenraad F van der SluijsTom van der Poll
Sep 13, 2003·Journal of Virology·Veronika von Messling, Roberto Cattaneo
Oct 15, 2003·Journal of Virology·Christel Schwegmann-WesselsGeorg Herrler
May 28, 2005·Journal of Virology·James Richard CarterRebecca Ellis Dutch
Feb 7, 2012·Journal of Virological Methods·Lieselot HouspieMarc Van Ranst
Oct 28, 2008·Biochemical and Biophysical Research Communications·Naokazu InoueMasaru Okabe
May 2, 2007·Annals of the New York Academy of Sciences·Sandra Diederich, Andrea Maisner
Dec 26, 2001·The Journal of General Virology·Helen W McL RixonRichard J Sugrue
Oct 5, 2013·The Journal of General Virology·Heather A LawlorRoderick S Tang
Mar 7, 2009·The Journal of General Virology·Christel Schwegmann-WeßelsGeorg Herrler
Jun 4, 2015·The Journal of Infectious Diseases·Nicholas J LennemannWendy Maury
Jun 8, 2018·Glycobiology·Victoria OrtegaHector C Aguilar
May 30, 2019·Medicina·Daifullah Al AboudAhmed S Abdel-Moneim
Apr 24, 2004·The Journal of General Virology·Wiebke KöhlGeorg Herrler
Mar 17, 2009·Bioorganic & Medicinal Chemistry·Danielle Skropeta

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