Analysis of filamentous process induction and nuclear localization properties of the HSV-2 serine/threonine kinase Us3

Virology. 2010 Feb 5;397(1):23-33. doi: 10.1016/j.virol.2009.11.012. Epub 2009 Nov 28.

Abstract

The Us3 serine/threonine kinase encoded by all alphaherpesviruses performs several important functions during virus multiplication. For example, expression of pseudorabies virus (PRV) Us3 causes reorganization of the actin cytoskeleton into filamentous processes (FPs) that promote cell-to-cell spread of virus infection. PRV Us3-induced FP formation requires Us3 kinase activity. To determine whether these characteristics were shared by HSV-2 Us3, expression plasmids for wild type (WT) and kinase dead (KD) Us3 variants were constructed. Expression of WT Us3 resulted in robust FP formation whereas expression of the KD Us3 variants did not. In the course of these experiments we noted that KD/K220 mutant Us3s were excluded from the nucleus in comparison to WT or KD/D305A Us3, prompting us to investigate Us3 nuclear shuttling properties. Herein we describe determinants of HSV-2 Us3-induced FP formation and present evidence for the presence of a leucine-rich nuclear export signal within HSV-2 Us3.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Substitution / genetics
  • Animals
  • Cell Nucleus / virology*
  • Chlorocebus aethiops
  • Herpesvirus 2, Human / physiology*
  • Humans
  • Mutation, Missense
  • Nuclear Export Signals
  • Protein Transport
  • Protein-Serine-Threonine Kinases / genetics
  • Protein-Serine-Threonine Kinases / metabolism*
  • Vero Cells
  • Viral Proteins / genetics
  • Viral Proteins / metabolism*
  • Virus Replication*

Substances

  • Nuclear Export Signals
  • Viral Proteins
  • Protein-Serine-Threonine Kinases
  • US3 protein, Human herpesvirus 2