Abstract
BackgroundThe visualization of viral proteins has been hindered by the resolution limit of conventional fluorescent microscopes, as the dimension of any single fluorescent signal is often greater than most virion particles. Super-resolution microscopy has the potential to unveil the distribution of proteins at the resolution approaching electron microscopy without relying on morphological features of existing characteristics of the biological specimen that are needed in EM.ResultsUsing direct stochastic optical reconstruction microscopy (dSTORM) to achieve a lateral resolution of 15–20 nm, we quantified the 2-D molecular distribution of the major structural proteins of the infectious human immunodeficiency virus type 1 (HIV-1) before and after infection of lymphoid cells. We determined that the HIV-1 matrix and capsid proteins undergo restructuring soon after HIV-1 infection.ConclusionsThis study provides the proof-of-concept for the use of dSTORM to visualize the changes in the molecular distribution of viral proteins during an infection.
Highlights
The visualization of viral proteins has been hindered by the resolution limit of conventional fluorescent microscopes, as the dimension of any single fluorescent signal is often greater than most virion particles
The molecular mechanisms of cell entry and replication are difficult to detect since the viral particle is smaller than the resolution limit of conventional fluorescent microscopes
This was confirmed by incubation of the target cells with an envelope-deficient human immunodeficiency virus type 1 (HIV-1), which was unable to enter the target cells and was cleaved by the pronase and no viral protein could be detected in these samples (Figure 1)
Summary
Using direct stochastic optical reconstruction microscopy (dSTORM) to achieve a lateral resolution of 15– 20 nm, we quantified the 2-D molecular distribution of the major structural proteins of the infectious human immunodeficiency virus type 1 (HIV-1) before and after infection of lymphoid cells. We determined that the HIV-1 matrix and capsid proteins undergo restructuring soon after HIV-1 infection
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