Microtubule plus end-associated CLIP-170 initiates HSV-1 retrograde transport in primary human cells.
Dynamic microtubules (MTs) continuously explore the intracellular environment and, through specialized plus end-tracking proteins (+TIPs), engage a variety of targets. However, the nature of cargoes that require +TIP-mediated capture for their movement on MTs remains poorly understood. Using RNA interference and dominant-negative approaches, combined with live cell imaging, we show that herpes simplex virus particles that have entered primary human cells exploit a +TIP complex comprising end-binding protein 1 (EB1), cytoplasmic linker protein 170 (CLIP-170), and dynactin-1 (DCTN1) to initiate retrograde transport. Depletion of these +TIPs completely blocked post-entry long-range transport of virus particles and suppressed infection ∼5,000-fold, whereas transferrin uptake, early endosome organization, and dynein-dependent movement of lysosomes and mitochondria remained unaffected. These findings provide the first insights into the earliest stages of viral engagement of MTs through specific +TIPs, akin to receptors, with therapeutic implications, and identify herpesvirus particles as one of a very limited number of cargoes absolutely dependent on CLIP-170-mediated capture to initiate transport in primary human cells.
Department of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611.
Vladimir
V
Jovasevic
Department of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 Department of Biochemistry and Molecular Biophysics, College of Physicians and Surgeons, Columbia University, New York, NY 10032 derek.walsh@northwestern.edu mojgan.naghavi@northwestern.edu.
Mojgan H
MH
Naghavi
Department of Microbiology-Immunology, Feinberg School of Medicine, Northwestern University, Chicago, IL 60611 Department of Microbiology, School of Medicine, New York University, New York, NY 10016 derek.walsh@northwestern.edu mojgan.naghavi@northwestern.edu.
Derek
D
Walsh
D000818:Animals; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D000072159:Dynactin Complex; D004398:Dyneins / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000378:metabolism*; D006801:Humans; D008247:Lysosomes / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics* / Q000378:metabolism*; D008870:Microtubules / Q000235:genetics / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D009363:Neoplasm Proteins / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering; D014168:Transferrin / Q000378:metabolism; D022161:Transport Vesicles / Q000378:metabolism; D014709:Vero Cells; D053586:Virus Internalization
C000606360:DCTN1 protein, human; D000072159:Dynactin Complex; C500985:MAPRE1 protein, human; D008869:Microtubule-Associated Proteins; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D014168:Transferrin; C076744:cytoplasmic linker protein 170; D004398:Dyneins