Dataset Information

Live Visualization of Hemagglutinin Dynamics During Infection by Using a Novel Reporter Influenza A Virus.

ABSTRACT: Live visualization of influenza A virus (IAV) structural proteins during viral infection in cells is highly sought objective to study different aspects of the viral replication cycle. To achieve this, we engineered an IAV to express a Tetra Cysteine tag (TC tag) from hemagglutinin (HA), which allows intracellular labeling of the engineered HA protein with biarsenic dyes and subsequent fluorescence detection. Using such constructs, we rescued a recombinant IAV with TC tag inserted in HA, in A/Puerto Rico/8/1934(H1N1) background (HA-TC). This recombinant HA-TC tag reporter IAV was replication-competent; however, as compared to wild type PR8 IAV, it was attenuated in multicycle replication. We confirmed expression of TC tag and biarsenical labeling of HA by immunofluorescence assay in cells infected with an HA-TC tag reporter IAV. Further, we used this reporter virus to visualize HA expression and translocation in IAV infected cells by live confocal imaging. We also tested the utility of the HA-TC IAV in testing chemical inhibitors of the HA translocation. Overall, HA-TC IAV is a versatile tool that will be useful for studying viral life cycle events, virus-host interactions, and anti-viral testing.

SUBMITTER: Dos Anjos Borges LG

PROVIDER: S-EPMC7354568 | biostudies-literature | 2020 Jun

REPOSITORIES: biostudies-literature

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Live Visualization of Hemagglutinin Dynamics During Infection by Using a Novel Reporter Influenza A Virus.

Dos Anjos Borges Luiz Gustavo LG Pisanelli Giuseppe G Khatun Oyahida O García-Sastre Adolfo A Tripathi Shashank S Tripathi Shashank S

Viruses 20200626 6

Live visualization of influenza A virus (IAV) structural proteins during viral infection in cells is highly sought objective to study different aspects of the viral replication cycle. To achieve this, we engineered an IAV to express a Tetra Cysteine tag (TC tag) from hemagglutinin (HA), which allows intracellular labeling of the engineered HA protein with biarsenic dyes and subsequent fluorescence detection. Using such constructs, we rescued a recombinant IAV with TC tag inserted in HA, in A/Pue ...[more]

PMID: 32604762

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Dataset Information

Live Visualization of Hemagglutinin Dynamics During Infection by Using a Novel Reporter Influenza A Virus.

Publications

Live Visualization of Hemagglutinin Dynamics During Infection by Using a Novel Reporter Influenza A Virus.

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