Application Manager, Functional Imaging. Business Unit Life Science & Applied Solutions, Leica Microsystems.
BIOGRAPHY
JUN 24, 2020 9:00 AM PDT
Advanced quantitative fluorescence microscopy to probe the molecular dynamics of viral entry.
Speaker
Event Date & Time
DATE: June 24, 2020
TIME: 9:00am PT, 12:00pm ET
Abstract
Some of the key factors in viral pathogenesis are the molecular mechanisms enabling viral entry into cells. Inhibitors of viral entry are often target of such mechanisms allowing to lower viral burdens during infections. Imaging experiments are specially well suited to probe the dynamic nature of viral entry. Here we will discuss how genetically-encoded biosensors paired with single virus tracking (SVT) can be used to assess the link between single cell metabolism and the corresponding ability of HIV particles to enter the cell (Coomer et al.). We will additionally see how quantitative functional imaging makes it possible to observe the time resolved stoichiometry of HIV receptor and coreceptor (CD4 and CCR5/CXCR4) during the formation of the HIV pre-fusion complex (Iliopoulou et al.). These insights into viral entry shed light on aspects of HIV entry that can be used as a basis to improve host-directed intervention strategies.
Coomer CA, Carlon-Andres I, Iliopoulou M, Dustin ML, Compeer EB, Compton AA, Padilla-Parra S. Single-cell glycolytic activity regulates membrane tension and HIV-1 fusion. PLoS Pathog. 2020 Feb 21;16(2):e1008359. doi: 10.1371/journal.ppat.1008359. eCollection 2020 Feb. PMID: 32084246
Iliopoulou M, Nolan R, Alvarez L, Watanabe Y, Coomer CA, Jakobsdottir GM, Bowden TA, Padilla-Parra S. A dynamic three-step mechanism drives the HIV-1 pre-fusion reaction. Nat Struct Mol Biol. 2018 Sep;25(9):814-822. doi: 10.1038/s41594-018-0113-x. Epub 2018 Aug 27. PMID: 30150645
Learning Objectives:
- Key aspects for quantitative fluorescence imaging together with single virus tracking
- How to multiplex biosensors to look at both viral fusion events and changes of single cell metabolism
- How to use genetically-encoded biosensors together with single virus tracking to gain insights into molecular mechanisms of viral entry
- How to use quantitative fluorescence imaging to decipher the time-resolved stoichiometry of membrane receptors during viral fusion
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