Phosphorylation of cytokeratin 17 by herpes simplex virus type 2 US3 protein kinase.
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.
Murata
Takayuki
T
Laboratory of Virology, Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Nagoya, Aichi 466-8550, Japan.
Goshima
Fumi
F
Nishizawa
Yuji
Y
Daikoku
Tohru
T
Takakuwa
Hiroki
H
Ohtsuka
Kenzo
K
Yoshikawa
Tetsushi
T
Nishiyama
Yukihiro
Y
D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D018258:Herpesvirus 2, Human / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D015151:Immunoblotting; D007382:Intermediate Filaments / Q000378:metabolism; D007633:Keratins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D010766:Phosphorylation; D011233:Precipitin Tests; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D025801:Ubiquitin / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis
D025801:Ubiquitin; D014764:Viral Proteins; D007633:Keratins; D017346:Protein Serine-Threonine Kinases; C084395:US3 protein, Human herpesvirus 2