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PMID: 1309913 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Alterations to influenza virus hemagglutinin cytoplasmic tail modulate virus infectivity.

Journal of virology ·Vol. 66 ·No. 2 ·1992-02-00 ·Pages 790-803

Simpson DA, Lamb RA

Abstract

The influenza virus hemagglutinin (HA) contains a cytoplasmic domain that consists of 10 to 11 amino acids, of which five residues have sequence identity for 10 of 13 HA subtypes. To investigate properties of these conserved residues, oligonucleotide-directed mutagenesis was performed, using an HA cDNA of influenza virus A/Udorn/72 (H3N2) to substitute the conserved cysteine residues with other residues, to delete the three C-terminal conserved residues, or to remove the entire cytoplasmic domain. The altered HAs were expressed in eukaryotic cells, and the rates of intracellular transport were examined. It was found that substitution of either conserved cysteine residue within the cytoplasmic domain did not affect the rate of intracellular transport, whereas deletion of residues within the C-terminal domain resulted in delayed cell surface expression. All the altered HAs were biologically active in hemadsorption and fusion assays. To investigate whether the wild-type HA and HAs with altered cytoplasmic tails could complement the influenza virus temperature-sensitive transport-defective HA mutant A/WSN/33 ts61S, the HA cDNAs were expressed by using a transient expression system and released virus was assayed by plaque analysis. The wild-type HA expression resulted in a release of approximately 10(3) PFU of virus per ml. Antibody neutralization of complemented virus indicated that the infectivity was due to incorporation of wild-type H3 HA into ts61S virions. Sucrose density gradient analysis of released virions showed that each of the HA cytoplasmic domain mutants was incorporated into virus particles. Virions containing HAs with substitution of the cysteine residues in the cytoplasmic domain were found to be infectious. However, no infectivity could be detected from virions containing HAs that had deletions in their cytoplasmic domains. Possible roles of the HA cytoplasmic domain in forming protein-protein interactions in virions and their involvement in the initiation of the infection process in cells are discussed.

MeSH Terms
Amino Acid Sequence Animals Cell Line Chickens Cytoplasm/physiology DNA, Viral/genetics Dimethyl Suberimidate Genetic Complementation Test Hemadsorption Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral/genetics Influenza A virus/genetics,physiology Kinetics Molecular Sequence Data Mutagenesis, Site-Directed Neutralization Tests RNA, Viral/genetics,isolation & purification Simian virus 40/genetics Viral Envelope Proteins/genetics Virion/genetics
Chemicals
DNA, Viral Hemagglutinin Glycoproteins, Influenza Virus Hemagglutinins, Viral RNA, Viral Viral Envelope Proteins Dimethyl Suberimidate
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Simpson D A
Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University, Evanston, Illinois 60208-3500.
Lamb R A
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.66.2.790-803.1992
Published
1992-02-00
Pages
790-803
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC240779
Subset
IM
Grants
NIAID NIH HHS · AI-20201 · United States
External Links
PubMed source DOI: 10.1128/JVI.66.2.790-803.1992 PMC Full Text View journal details
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