Bicaudal D2 Facilitates the Cytoplasmic Trafficking and Nuclear Import of HIV-1 Genomes During Infection Ed Campbell Microbiology and Immunology Loyola.

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Presentation transcript:

Bicaudal D2 Facilitates the Cytoplasmic Trafficking and Nuclear Import of HIV-1 Genomes During Infection Ed Campbell Microbiology and Immunology Loyola University, Chicago

HIV-1 Trafficks Along Microtubules Vpr Tubulin McDonald et al., JCB 2002

Bicaudal D Homolog 2 Protein (BICD2) Cargo Binding Domain Jha et al. Biochemical Society Transactions 2015

Bicaudal D Homolog 2 Protein (BICD2) Cargo Binding Domain Jha et al. Biochemical Society Transactions 2015

CRISPR knockout BICD2 Cell Lines

HIV-1 Infectivity is Impaired in BICD2 knockout cells THP1 differentiated macrophages TZM-bl

Viral Fusion is Unaffected in BICD2 knockout Cells Blam-Vpr Viral Fusion Assay

Reverse Transcription and Nuclear Import 24h post Infection

Reverse Transcription and Nuclear Import 24h post Infection

Is BICD2 the dynein cargo adaptor for HIV-1 Does BICD2 Interact with HIV-1 capsid? Is BICD2 required for viral trafficking?

BICD2 Binds Capsid Through the Cargo Binding CC3 Domain BICD2-CC2-CC3-HA BICD2-CC2-CC3-HA

BICD2 Required for Efficient Cytoplasmic Trafficking of HIV-1 Viral Particles BICD2 KO TZM-Bl Cells TZM-Bl Cells

BICD2 Required for Efficient Cytoplasmic Trafficking of HIV-1 Viral Particles Fused unfused Fused unfused ns *** ns ***

Depletion of BICD2 prevents Directed Motion of Viral Particles Control Cells MSD (µM2) Δt (sec) R2=0.9977 Xiao et al., JVI 2012

Depletion of BICD2 Results in Induction of Several ISGs upon Viral infection THP1 Cells 24h post Infection

Summary Microtubules Nucleus Plasma membrane Cytoplasm CD4 Coreceptors Plasma membrane Cytoplasm Microtubules Dynein-dynactin Complex BICD2 Nuclear Pore Nucleus

Summary Microtubules Nucleus Plasma membrane Cytoplasm CD4 Coreceptors Plasma membrane Cytoplasm Microtubules Dynein-dynactin Complex BICD2 Nuclear Pore Nucleus

Summary Microtubules Nucleus Plasma membrane Cytoplasm CD4 Microtubules Coreceptors Plasma membrane Nuclear Pore Nucleus Cytoplasm Dynein-dynactin Complex BICD2

Acknowledgements Adarsh Dharan Campbell Lab Omar Abdel Rahim Sabrina Imam Sarah Talley Sevnur Komurlu Bill Flavin Nikolai Smolin Alexander Simon Grace Ispas Kevin Burbidge Benjamin Cook Stephanie Zack Virginia Zwikelmaier Albert Einstein College of Medicine Felipe Diaz-Griffero Silvana Opp Northwestern Universty Hope Lab Utrecht University Casper Hoogenraad NIH R01 AI120956

BICD2 Co-Traffics with HIV-1 Viral Particles GFP-BICD2 JRFLg-R7/GIR