Evaluation of Glu11 and Gly8 of the H5N1 influenza hemagglutinin fusion peptide in membrane fusion using pseudotype virus and reverse genetics

Y. Su, Xingguo Zhu, Y. Wang, M. Wu, P. Tien

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Influenza viruses gain entry into host cells by binding to cellular receptors and promoting the fusion of the viral envelope with the host cell membrane. The fusion peptide of influenza hemagglutinin (HA) is crucial for fusion. To examine the structural and functional roles of amino acids E11 and G8 of the H5 HA fusion peptide, a series of fusion mutants was generated. We determined the effect of each mutation on fusion activity and infection of rescued recombinant virus by polykaryon formation, cell-cell fusion assay, HA pseudovirus transduction and reverse genetics. Our findings indicate that E11V and E11A mutants dramatically inhibit fusion and that at position 11 a polar residue such as glutamic acid or serine may be desirable for preserving the fusion activity. More interestingly, one mutation (G8E) raised the threshold pH of polykaryon formation. Our results suggest that G8 as well as E11 play an important functional and structural role in membrane fusion and that the polarity of E11 is crucial for fusion activity. Finally, we developed an assay based on a reporter gene plus pseudotyped virus that could sensitively detect fusion activity.

Original languageEnglish (US)
Pages (from-to)247-257
Number of pages11
JournalArchives of Virology
Volume153
Issue number2
DOIs
StatePublished - Feb 1 2008

Fingerprint

Reverse Genetics
Membrane Fusion
Hemagglutinins
Human Influenza
Viruses
Virus Internalization
Peptides
Mutation
Cell Fusion
Orthomyxoviridae
Reporter Genes
Serine
Glutamic Acid
Cell Membrane
Amino Acids
Infection
avian influenza A virus hemagglutinin

ASJC Scopus subject areas

  • Virology

Cite this

Evaluation of Glu11 and Gly8 of the H5N1 influenza hemagglutinin fusion peptide in membrane fusion using pseudotype virus and reverse genetics. / Su, Y.; Zhu, Xingguo; Wang, Y.; Wu, M.; Tien, P.

In: Archives of Virology, Vol. 153, No. 2, 01.02.2008, p. 247-257.

Research output: Contribution to journalArticle

@article{d552f1bd75664d94bfbce69c5fad58f0,
title = "Evaluation of Glu11 and Gly8 of the H5N1 influenza hemagglutinin fusion peptide in membrane fusion using pseudotype virus and reverse genetics",
abstract = "Influenza viruses gain entry into host cells by binding to cellular receptors and promoting the fusion of the viral envelope with the host cell membrane. The fusion peptide of influenza hemagglutinin (HA) is crucial for fusion. To examine the structural and functional roles of amino acids E11 and G8 of the H5 HA fusion peptide, a series of fusion mutants was generated. We determined the effect of each mutation on fusion activity and infection of rescued recombinant virus by polykaryon formation, cell-cell fusion assay, HA pseudovirus transduction and reverse genetics. Our findings indicate that E11V and E11A mutants dramatically inhibit fusion and that at position 11 a polar residue such as glutamic acid or serine may be desirable for preserving the fusion activity. More interestingly, one mutation (G8E) raised the threshold pH of polykaryon formation. Our results suggest that G8 as well as E11 play an important functional and structural role in membrane fusion and that the polarity of E11 is crucial for fusion activity. Finally, we developed an assay based on a reporter gene plus pseudotyped virus that could sensitively detect fusion activity.",
author = "Y. Su and Xingguo Zhu and Y. Wang and M. Wu and P. Tien",
year = "2008",
month = "2",
day = "1",
doi = "10.1007/s00705-007-1088-9",
language = "English (US)",
volume = "153",
pages = "247--257",
journal = "Archives of Virology",
issn = "0304-8608",
publisher = "Springer Wien",
number = "2",

}

TY - JOUR

T1 - Evaluation of Glu11 and Gly8 of the H5N1 influenza hemagglutinin fusion peptide in membrane fusion using pseudotype virus and reverse genetics

AU - Su, Y.

AU - Zhu, Xingguo

AU - Wang, Y.

AU - Wu, M.

AU - Tien, P.

PY - 2008/2/1

Y1 - 2008/2/1

N2 - Influenza viruses gain entry into host cells by binding to cellular receptors and promoting the fusion of the viral envelope with the host cell membrane. The fusion peptide of influenza hemagglutinin (HA) is crucial for fusion. To examine the structural and functional roles of amino acids E11 and G8 of the H5 HA fusion peptide, a series of fusion mutants was generated. We determined the effect of each mutation on fusion activity and infection of rescued recombinant virus by polykaryon formation, cell-cell fusion assay, HA pseudovirus transduction and reverse genetics. Our findings indicate that E11V and E11A mutants dramatically inhibit fusion and that at position 11 a polar residue such as glutamic acid or serine may be desirable for preserving the fusion activity. More interestingly, one mutation (G8E) raised the threshold pH of polykaryon formation. Our results suggest that G8 as well as E11 play an important functional and structural role in membrane fusion and that the polarity of E11 is crucial for fusion activity. Finally, we developed an assay based on a reporter gene plus pseudotyped virus that could sensitively detect fusion activity.

AB - Influenza viruses gain entry into host cells by binding to cellular receptors and promoting the fusion of the viral envelope with the host cell membrane. The fusion peptide of influenza hemagglutinin (HA) is crucial for fusion. To examine the structural and functional roles of amino acids E11 and G8 of the H5 HA fusion peptide, a series of fusion mutants was generated. We determined the effect of each mutation on fusion activity and infection of rescued recombinant virus by polykaryon formation, cell-cell fusion assay, HA pseudovirus transduction and reverse genetics. Our findings indicate that E11V and E11A mutants dramatically inhibit fusion and that at position 11 a polar residue such as glutamic acid or serine may be desirable for preserving the fusion activity. More interestingly, one mutation (G8E) raised the threshold pH of polykaryon formation. Our results suggest that G8 as well as E11 play an important functional and structural role in membrane fusion and that the polarity of E11 is crucial for fusion activity. Finally, we developed an assay based on a reporter gene plus pseudotyped virus that could sensitively detect fusion activity.

UR - http://www.scopus.com/inward/record.url?scp=39149129436&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=39149129436&partnerID=8YFLogxK

U2 - 10.1007/s00705-007-1088-9

DO - 10.1007/s00705-007-1088-9

M3 - Article

C2 - 18030546

AN - SCOPUS:39149129436

VL - 153

SP - 247

EP - 257

JO - Archives of Virology

JF - Archives of Virology

SN - 0304-8608

IS - 2

ER -