title: Nuclear herpesvirus capsid motility is not dependent on F-actin.
abstract: A considerable part of the herpesvirus life cycle takes place in the host nucleus. While much progress has been made to understand the molecular processes required for virus replication in the nucleus, much less is known about the temporal and spatial dynamics of these events. Previous studies have suggested that nuclear capsid motility is directed and dependent on actin filaments (F-actin), possibly using a myosin-based, ATP-dependent mechanism. However, the conclusions from these studies were indirect. They either relied on the effects of F-actin depolymerizing drugs to deduce an F-actin dependency or they visualized nuclear F-actin but failed to show a direct link to capsid motility. Moreover, no direct link between nuclear capsid motility and a molecular motor has been established. In this report, we reinvestigate the involvement of F-actin in nuclear herpesvirus capsid transport. We show for representative members of all three herpesvirus subfamilies that nuclear capsid motility is not dependent on nuclear F-actin and that herpesvirus infection does not induce nuclear F-actin in primary fibroblasts. Moreover, in these cells, three F-actin-inhibiting drugs failed to effect capsid motility. Only latrunculin A treatment stalled nuclear capsids but did so by an unexpected effect: the drug induced actin rods in the nucleus. Immobile capsids accumulated around actin rods, and immunoprecipitation experiments suggested that capsid motility stopped because latrunculin-induced actin rods nonspecifically bind nuclear capsids. Interestingly, capsid motility was unaffected in cells that do not induce actin rods. Based on these data, we conclude that herpesvirus nuclear capsid motility is not dependent on F-actin. Importance: Herpesviruses are large DNA viruses whose replication is dependent on the host nucleus. However, we do not understand how key nuclear processes, including capsid assembly, genome replication, capsid packaging, and nuclear egress, are dynamically connected in space and time. Fluorescence live-cell microscopy revealed that nuclear capsids are highly mobile early in infection. Two studies suggested that this motility might be due to active myosin-based transport of capsids on nuclear F-actin. However, direct evidence for such motor-based transport is lacking. We revisited this phenomenon and found no evidence that nuclear capsid motility depended on F-actin. Our results reopen the question of how nuclear herpesvirus capsids move in the host nucleus.
authors.forename: Jens B
authors.initials: JB
authors.lastname: Bosse
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, New Jersey, USA Princeton Neuroscience Institute, Princeton University, Princeton, New Jersey, USA.
authors.forename: Stina
authors.initials: S
authors.lastname: Virding
authors.affiliation: Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA.
authors.forename: Stephan Y
authors.initials: SY
authors.lastname: Thiberge
authors.forename: Julian
authors.initials: J
authors.lastname: Scherer
authors.affiliation: Microbiologie Fondamental et Pathogénicité, CNRS UMR 5234, University of Bordeaux, Segalen, Bordeaux, France.
authors.forename: Harald
authors.initials: H
authors.lastname: Wodrich
authors.affiliation: Max von Pettenkofer-Institut, Ludwig-Maximilians-Universität, Munich, Germany.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Ruzsics
authors.affiliation: Max von Pettenkofer-Institut, Ludwig-Maximilians-Universität, Munich, Germany.
authors.forename: Ulrich H
authors.initials: UH
authors.lastname: Koszinowski
authors.affiliation: lenquist@princeton.edu.
authors.forename: Lynn W
authors.initials: LW
authors.lastname: Enquist
mesh_terms: D000199:Actins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000818:Animals; D001692:Biological Transport; D002213:Capsid / Q000378:metabolism*; D002467:Cell Nucleus / Q000821:virology*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000821:virology; D006564:Herpesviridae / Q000502:physiology*; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence
chemical_list: D000199:Actins