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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 on theme: "Bicaudal D2 Facilitates the Cytoplasmic Trafficking and Nuclear Import of HIV-1 Genomes During Infection Ed Campbell Microbiology and Immunology Loyola."— Presentation transcript:

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

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

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

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

5 CRISPR knockout BICD2 Cell Lines

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

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

8 Reverse Transcription and Nuclear Import
24h post Infection

9 Reverse Transcription and Nuclear Import
24h post Infection

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

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

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

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

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

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

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

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

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

19 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 

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

21

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