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HIV-1 capsids bind and exploit the kinesin-1 adaptor FEZ1 for inward movement to the nucleus.

Malikov V, da Silva ES, Jovasevic V, Bennett G, de Souza Aranha Vieira DA, Schulte B, Diaz-Griffero F, Walsh D, Naghavi MH - Nat Commun (2015)

Bottom Line: Furthermore, both dynein and kinesin-1 motors are required for HIV-1 trafficking to the nucleus.Finally, the ability of exogenously expressed FEZ1 to promote early HIV-1 infection requires binding to kinesin-1.Our findings demonstrate that opposing motors both contribute to early HIV-1 movement and identify the kinesin-1 adaptor, FEZ1 as a capsid-associated host regulator of this process usurped by HIV-1 to accomplish net inward movement towards the nucleus.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Microbiology-Immunology, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA [2] Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York 10032, USA.

ABSTRACT
Intracellular transport of cargos, including many viruses, involves directed movement on microtubules mediated by motor proteins. Although a number of viruses bind motors of opposing directionality, how they associate with and control these motors to accomplish directed movement remains poorly understood. Here we show that human immunodeficiency virus type 1 (HIV-1) associates with the kinesin-1 adaptor protein, Fasiculation and Elongation Factor zeta 1 (FEZ1). RNAi-mediated FEZ1 depletion blocks early infection, with virus particles exhibiting bi-directional motility but no net movement to the nucleus. Furthermore, both dynein and kinesin-1 motors are required for HIV-1 trafficking to the nucleus. Finally, the ability of exogenously expressed FEZ1 to promote early HIV-1 infection requires binding to kinesin-1. Our findings demonstrate that opposing motors both contribute to early HIV-1 movement and identify the kinesin-1 adaptor, FEZ1 as a capsid-associated host regulator of this process usurped by HIV-1 to accomplish net inward movement towards the nucleus.

No MeSH data available.


Related in: MedlinePlus

Kinesin-1 depletion does not affect dynein, an inward motor required for HIV-1 infection(a–b) Effects of kinesin-1 depletion on dynein levels and distribution. NHDF cells were treated with control siRNA (Ctrl) or siRNAs targeting Kif5A or Kif5B. 48h post-transfection cells were either lysed and analyzed by WB (a) or fixed and stained (b) using antibodies to kinesin-1 (Kif5A/B), Kif5B, dynein, trans-golgi network marker (TNG46) or β-actin. Molecular weight markers (in kDa) are shown to the right of WBs. Scale bars represent 10 μm. (c–e) Dynein is required for early HIV-1 infection. NHDF (c), CHME3 (d) or Jurkat (e) cells were treated with the indicated concentrations of the dynein inhibitor Ciliobrevin D for 30 min followed by HIV-1-VSV-luc infection in the presence of inhibitor. Luciferase activity was measured 48h post-infection to determine levels of infection. Error bars represent standard deviation from three independent experiments.
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Figure 5: Kinesin-1 depletion does not affect dynein, an inward motor required for HIV-1 infection(a–b) Effects of kinesin-1 depletion on dynein levels and distribution. NHDF cells were treated with control siRNA (Ctrl) or siRNAs targeting Kif5A or Kif5B. 48h post-transfection cells were either lysed and analyzed by WB (a) or fixed and stained (b) using antibodies to kinesin-1 (Kif5A/B), Kif5B, dynein, trans-golgi network marker (TNG46) or β-actin. Molecular weight markers (in kDa) are shown to the right of WBs. Scale bars represent 10 μm. (c–e) Dynein is required for early HIV-1 infection. NHDF (c), CHME3 (d) or Jurkat (e) cells were treated with the indicated concentrations of the dynein inhibitor Ciliobrevin D for 30 min followed by HIV-1-VSV-luc infection in the presence of inhibitor. Luciferase activity was measured 48h post-infection to determine levels of infection. Error bars represent standard deviation from three independent experiments.

Mentions: It has been shown previously that in some cellular contexts the inhibition of kinesin-1 activity can result in the accumulation of dynein at the trans-golgi network (TGN)25,26. This might also explain the block to early HIV-1 infection. To test this, the levels and distribution of dynein in cells treated with control, Kif5A or Kif5B siRNAs were examined. WB analysis revealed that Kif5A or Kif5B depletion did not affect the levels of dynein or β-actin compared to control samples (Fig. 5a). Furthermore, immunofluorescence (IF) analysis of fixed cells stained using antibodies against Kif5, dynein intermediate chain (IC74) and the TGN marker, TGN46 revealed no notable differences in the distribution of dynein between cells treated with control or kinesin-1-specific siRNAs, nor were increases in dynein accumulation at the TGN detected (Fig. 5b). These findings demonstrated that the block to HIV-1 infection in kinesin-1-depleted cells was not due to defects in dynein levels or localization.


HIV-1 capsids bind and exploit the kinesin-1 adaptor FEZ1 for inward movement to the nucleus.

Malikov V, da Silva ES, Jovasevic V, Bennett G, de Souza Aranha Vieira DA, Schulte B, Diaz-Griffero F, Walsh D, Naghavi MH - Nat Commun (2015)

Kinesin-1 depletion does not affect dynein, an inward motor required for HIV-1 infection(a–b) Effects of kinesin-1 depletion on dynein levels and distribution. NHDF cells were treated with control siRNA (Ctrl) or siRNAs targeting Kif5A or Kif5B. 48h post-transfection cells were either lysed and analyzed by WB (a) or fixed and stained (b) using antibodies to kinesin-1 (Kif5A/B), Kif5B, dynein, trans-golgi network marker (TNG46) or β-actin. Molecular weight markers (in kDa) are shown to the right of WBs. Scale bars represent 10 μm. (c–e) Dynein is required for early HIV-1 infection. NHDF (c), CHME3 (d) or Jurkat (e) cells were treated with the indicated concentrations of the dynein inhibitor Ciliobrevin D for 30 min followed by HIV-1-VSV-luc infection in the presence of inhibitor. Luciferase activity was measured 48h post-infection to determine levels of infection. Error bars represent standard deviation from three independent experiments.
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Related In: Results  -  Collection

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Figure 5: Kinesin-1 depletion does not affect dynein, an inward motor required for HIV-1 infection(a–b) Effects of kinesin-1 depletion on dynein levels and distribution. NHDF cells were treated with control siRNA (Ctrl) or siRNAs targeting Kif5A or Kif5B. 48h post-transfection cells were either lysed and analyzed by WB (a) or fixed and stained (b) using antibodies to kinesin-1 (Kif5A/B), Kif5B, dynein, trans-golgi network marker (TNG46) or β-actin. Molecular weight markers (in kDa) are shown to the right of WBs. Scale bars represent 10 μm. (c–e) Dynein is required for early HIV-1 infection. NHDF (c), CHME3 (d) or Jurkat (e) cells were treated with the indicated concentrations of the dynein inhibitor Ciliobrevin D for 30 min followed by HIV-1-VSV-luc infection in the presence of inhibitor. Luciferase activity was measured 48h post-infection to determine levels of infection. Error bars represent standard deviation from three independent experiments.
Mentions: It has been shown previously that in some cellular contexts the inhibition of kinesin-1 activity can result in the accumulation of dynein at the trans-golgi network (TGN)25,26. This might also explain the block to early HIV-1 infection. To test this, the levels and distribution of dynein in cells treated with control, Kif5A or Kif5B siRNAs were examined. WB analysis revealed that Kif5A or Kif5B depletion did not affect the levels of dynein or β-actin compared to control samples (Fig. 5a). Furthermore, immunofluorescence (IF) analysis of fixed cells stained using antibodies against Kif5, dynein intermediate chain (IC74) and the TGN marker, TGN46 revealed no notable differences in the distribution of dynein between cells treated with control or kinesin-1-specific siRNAs, nor were increases in dynein accumulation at the TGN detected (Fig. 5b). These findings demonstrated that the block to HIV-1 infection in kinesin-1-depleted cells was not due to defects in dynein levels or localization.

Bottom Line: Furthermore, both dynein and kinesin-1 motors are required for HIV-1 trafficking to the nucleus.Finally, the ability of exogenously expressed FEZ1 to promote early HIV-1 infection requires binding to kinesin-1.Our findings demonstrate that opposing motors both contribute to early HIV-1 movement and identify the kinesin-1 adaptor, FEZ1 as a capsid-associated host regulator of this process usurped by HIV-1 to accomplish net inward movement towards the nucleus.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Microbiology-Immunology, Northwestern University Feinberg School of Medicine, Chicago, Illinois 60611, USA [2] Department of Biochemistry and Molecular Biophysics, Columbia University, New York, New York 10032, USA.

ABSTRACT
Intracellular transport of cargos, including many viruses, involves directed movement on microtubules mediated by motor proteins. Although a number of viruses bind motors of opposing directionality, how they associate with and control these motors to accomplish directed movement remains poorly understood. Here we show that human immunodeficiency virus type 1 (HIV-1) associates with the kinesin-1 adaptor protein, Fasiculation and Elongation Factor zeta 1 (FEZ1). RNAi-mediated FEZ1 depletion blocks early infection, with virus particles exhibiting bi-directional motility but no net movement to the nucleus. Furthermore, both dynein and kinesin-1 motors are required for HIV-1 trafficking to the nucleus. Finally, the ability of exogenously expressed FEZ1 to promote early HIV-1 infection requires binding to kinesin-1. Our findings demonstrate that opposing motors both contribute to early HIV-1 movement and identify the kinesin-1 adaptor, FEZ1 as a capsid-associated host regulator of this process usurped by HIV-1 to accomplish net inward movement towards the nucleus.

No MeSH data available.


Related in: MedlinePlus