Phosphorylation of cytokeratin 17 by herpes simplex virus type 2 US3 protein kinase

Microbiol Immunol. 2002;46(10):707-19. doi: 10.1111/j.1348-0421.2002.tb02755.x.

Abstract

We previously reported the establishment of an HEp2 cell line which expresses the US3 protein kinase (PK) of herpes simplex virus type 2 (HSV-2) upon induction with IPTG. Here we report that expression, phosphorylation and ubiquitination of cytokeratin 17 (CK17) are enhanced in US3-expressing HEp2 cells. In vitro kinase and co-immunoprecipitation assays provided evidence that US3 PK directly phosphorylates CK17. Expression of US3 PK caused a significant decrease in filamentous staining of CK17, suggesting that phosphorylation of CK17 by US3 PK causes a disruption of intermediate filaments. Our observations suggest a role for US3 in the regulation of CKs and intermediate filaments in cells. Moreover, we found that infection of a keratinocyte-derived cell line, A431, with a US3-deficient virus, results in cytopathic effects that are morphologically distinct from those induced by wild-type and revertant viruses, suggesting that US3 PK may be important for interaction between HSV-2 and peripheral epithelial cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Electrophoresis, Gel, Two-Dimensional
  • Herpesvirus 2, Human / enzymology*
  • Herpesvirus 2, Human / genetics
  • Humans
  • Immunoblotting
  • Intermediate Filaments / metabolism
  • Keratins / metabolism*
  • Microscopy, Fluorescence
  • Phosphorylation
  • Precipitin Tests
  • Protein-Serine-Threonine Kinases / metabolism*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
  • Ubiquitin / metabolism
  • Viral Proteins / biosynthesis

Substances

  • Ubiquitin
  • Viral Proteins
  • Keratins
  • Protein-Serine-Threonine Kinases
  • US3 protein, Human herpesvirus 2