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Quantitatively Dissecting Triple Roles of Dynactin in Dynein-Driven Transport of Influenza Virus by Quantum Dot-Based
Dan-Dan Fu1, Li-Juan Zhang1, Bo Tang1
1College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, P.R. China.
Abstract:
After entering host cells by endocytosis, influenza A virus (IAV) is transported along microfilaments and then transported by dynein along microtubules (MTs) to the perinuclear region for genome release. Understanding the mechanisms of dynein-driven transport is significant for a comprehensive understanding of IAV infection. In this work, the roles of dynactin in dynein-driven transport of IAV were quantitatively dissected in situ using quantum dot-based single-virus tracking. It was revealed that dynactin was essential for dynein to transport IAV toward the nucleus. After virus entry, virus-carrying vesicles bound to dynein and dynactin before being delivered to MTs. The attachment of dynein to the vesicles was dependent on dynactin and its subunits, p150Glued and Arp1. Once viruses reached MTs, dynactin-assisted dynein initiates retrograde transport of IAV. Importantly, the retrograde transport of viruses could be initiated at both plus ends (32%) and other regions on MTs (68%). Subsequently, dynactin accompanied and assisted dynein to persistently transport the virus along MTs in the retrograde direction. This study revealed the dynactin-dependent dynein-driven transport process of IAV, enhancing our understanding of IAV infection and providing important insights into the cell's endocytic transport mechanism.
Insights
Dynactin is essential for dynein to transport influenza A virus (IAV) within host cells. This study reveals dynactin
Area of Science:
- Cell Biology
- Virology
- Molecular Motor Function
Background:
- Influenza A virus (IAV) infection requires intracellular transport for genome release.
- Dynein motor proteins are crucial for moving IAV towards the nucleus after cell entry.
- Understanding the precise mechanisms of dynein-mediated IAV transport is vital for comprehending viral infection cycles.
Purpose of the Study:
- To quantitatively investigate the role of dynactin in dynein-driven transport of IAV within host cells.
- To elucidate the molecular mechanisms by which dynactin influences dynein's interaction with IAV-containing vesicles.
- To determine the specific contributions of dynactin and its subunits to the retrograde transport of IAV along microtubules.
Main Methods:
- Utilized quantum dot-based single-virus tracking for in situ analysis of IAV intracellular movement.
- Quantitatively assessed the dependence of dynein-vesicle attachment on dynactin and its subunits (p150Glued, Arp1).
- Tracked virus transport dynamics along microtubules, noting initiation sites and persistent movement.
Main Results:
- Dynactin is indispensable for dynein-mediated transport of IAV towards the perinuclear region.
- Dynein attaches to IAV-carrying vesicles in a dynactin-dependent manner, involving subunits p150Glued and Arp1.
- Dynactin-assisted dynein initiates retrograde transport of IAV along microtubules, with transport initiation occurring at both microtubule plus ends and other regions.
- Dynactin actively accompanies and facilitates dynein's sustained retrograde movement of IAV along microtubules.
Conclusions:
- Dynactin plays a critical, multifaceted role in dynein-driven intracellular transport of influenza A virus.
- This study reveals a dynactin-dependent mechanism for dynein-mediated IAV retrograde transport, enhancing understanding of viral infection.
- Provides key insights into the fundamental cellular machinery governing endocytic transport processes.
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