Intercellular trafficking and cytotoxicity of recombinant HSV-1 thymidine kinase fused with HSV-2 US11 RXP repeat peptide.
To improve the therapeutic efficacy of herpes simplex virus type 1 (HSV-1) thymidine kinase (tk)/ganciclovir (GCV) therapy, we have made recombinant tk chimeras fused with the arginine-rich (RXP) repeat of herpes simplex virus type 2 (HSV-2) US11 and examined their activity of intercellular trafficking and cytotoxicity. When examined the immunofluorescence staining patterns of RXP/tk fusion proteins in transfected COS7 cells, the RXP chimeras revealed a conservation of the trafficking activity of RXP. We also found that transfection of tkC Delta 6-RXP (lacking the C-terminal six amino residues of tk), tk-RXP, and tkN Delta 66-RXP (lacking the N-terminal 66 amino residues of tk) induced apoptosis even in the absence of GCV. The results suggest that these tk/RXP chimeras themselves have apoptosis-inducing activity, and that the HSV tk nucleoside-binding domain may be involved in the induction of apoptosis. Furthermore, treatment with 5 muM GCV induced efficient cell death in cells tranfected with tk-RXP in comparison to the cells transfected with tk (P < 0.0001).
Luo
Chenhong
C
Department of Virology, Nagoya University Graduate School of Medicine, Tsurumai-cho 65, showa-ku, 466-8550, Nagoya, Japan.
Nawa
Akihiro
A
Yamauchi
Youhei
Y
Kohno
Shinichi
S
Ushijima
Youko
Y
Goshima
Fumi
F
Kikkawa
Fumitaka
F
Nishiyama
Yukihiro
Y
D000818:Animals; D001120:Arginine / Q000737:chemistry / Q000235:genetics; D019556:COS Cells; D016923:Cell Death / Q000187:drug effects; D002522:Chlorocebus aethiops; D015774:Ganciclovir / Q000494:pharmacology; D041722:Genes, Transgenic, Suicide; D018259:Herpesvirus 1, Human / Q000201:enzymology / Q000235:genetics; D018258:Herpesvirus 2, Human / Q000235:genetics; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism; D011392:Proline / Q000737:chemistry / Q000235:genetics; D021381:Protein Transport; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*; D012091:Repetitive Sequences, Nucleic Acid; D013937:Thymidine Kinase / Q000235:genetics / Q000378:metabolism*; D014018:Tissue Distribution; D014162:Transfection; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D010446:Peptide Fragments; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C074546:US11 protein, herpesvirus; D014764:Viral Proteins; D001120:Arginine; D011392:Proline; D013937:Thymidine Kinase; D015774:Ganciclovir