Inner tegument proteins of Herpes Simplex Virus are sufficient for intracellular capsid motility in neurons but not for axonal targeting

• Published in: PLoS pathogens Vol. 13; no. 12; p. e1006813
• Main Authors: Buch, Anna, Müller, Oliver, Ivanova, Lyudmila, Döhner, Katinka, Bialy, Dagmara, Bosse, Jens B, Pohlmann, Anja, Binz, Anne, Hegemann, Maike, Nagel, Claus-Henning, Koltzenburg, Martin, Viejo-Borbolla, Abel, Rosenhahn, Bodo, Bauerfeind, Rudolf, Sodeik, Beate
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 01.12.2017; Public Library of Science (PLoS)
• Subjects: Animals; Axonal Transport; Axons - ultrastructure; Axons - virology; Biology and Life Sciences; Capsid - physiology; Capsid - ultrastructure; Cells, Cultured; Chlorocebus aethiops; Ganglia, Spinal - virology; Herpes Simplex - virology; Herpesvirus 1, Human - genetics; Herpesvirus 1, Human - pathogenicity; Herpesvirus 1, Human - physiology; Host-Pathogen Interactions; Humans; Mice; Microscopy, Electron, Transmission; Movement - physiology; Mutation; Neurons - ultrastructure; Neurons - virology; Vero Cells; Viral Proteins - genetics; Viral Proteins - physiology; Viral Structural Proteins - genetics; Viral Structural Proteins - physiology
• ISSN: 1553-7374; 1553-7366; 1553-7374
• DOI: 10.1371/journal.ppat.1006813

Upon reactivation from latency and during lytic infections in neurons, alphaherpesviruses assemble cytosolic capsids, capsids associated with enveloping membranes, and transport vesicles harboring fully enveloped capsids. It is debated whether capsid envelopment of herpes simplex virus (HSV) is completed in the soma prior to axonal targeting or later, and whether the mechanisms are the same in neurons derived from embryos or from adult hosts. We used HSV mutants impaired in capsid envelopment to test whether the inner tegument proteins pUL36 or pUL37 necessary for microtubule-mediated capsid transport were sufficient for axonal capsid targeting in neurons derived from the dorsal root ganglia of adult mice. Such neurons were infected with HSV1-ΔUL20 whose capsids recruited pUL36 and pUL37, with HSV1-ΔUL37 whose capsids associate only with pUL36, or with HSV1-ΔUL36 that assembles capsids lacking both proteins. While capsids of HSV1-ΔUL20 were actively transported along microtubules in epithelial cells and in the somata of neurons, those of HSV1-ΔUL36 and -ΔUL37 could only diffuse in the cytoplasm. Employing a novel image analysis algorithm to quantify capsid targeting to axons, we show that only a few capsids of HSV1-ΔUL20 entered axons, while vesicles transporting gD utilized axonal transport efficiently and independently of pUL36, pUL37, or pUL20. Our data indicate that capsid motility in the somata of neurons mediated by pUL36 and pUL37 does not suffice for targeting capsids to axons, and suggest that capsid envelopment needs to be completed in the soma prior to targeting of herpes simplex virus to the axons, and to spreading from neurons to neighboring cells.