ePrints@IIScePrints@IISc Home | About | Browse | Latest Additions | Advanced Search | Contact | Help

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

Dos Anjos Borges, LG and Pisanelli, G and Khatun, O and García-Sastre, A and Tripathi, S (2020) Live Visualization of Hemagglutinin Dynamics during Infection by Using a Novel Reporter Influenza A Virus. In: Viruses, 12 (6).

vir_12-06_2020.pdf - Published Version

Download (2MB) | Preview
Official URL: https://dx.doi.org/10.3390/v12060687


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 ofHAby 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. © 2020 by the authors.

Item Type: Journal Article
Publication: Viruses
Publisher: MDPI AG
Additional Information: The copyright of this article belongs to MDPI AG
Department/Centre: Division of Biological Sciences > Microbiology & Cell Biology
Division of Biological Sciences > Centre for Infectious Disease Research
Date Deposited: 24 Aug 2020 11:14
Last Modified: 24 Aug 2020 11:14
URI: http://eprints.iisc.ac.in/id/eprint/66047

Actions (login required)

View Item View Item