Herpes simplex virus type 1 (HSV-1) establishes life-long latent infection in sensory neurones, making it an ideal gene therapy vector to target neuronal cells. To study retrograde transport of HSV-1, capsid and tegument proteins were tested for interactions with the motor cytoplasmic dynein, using a yeast two-hybrid system and in vitro pull down assays. A strong interaction was demonstrated between the HSV-1 outer capsid protein VP26 (UL35) and the homologous 14 kDa dynein light chains RP3 and Tctex1. The functional importance of VP26 for retrograde transport was confirmed by microinjecting recombinant HSV-1 capsids into living cells. Capsids containing VP26 moved closer to the cell nucleus, while capsids without VP26 remained in a random distribution. Our results suggest that the HSV-1 outer capsid protein VP26 mediates binding of incoming capsids to the retrograde motor cytoplasmic dynein during cellular infection, through interactions with dynein light chains. It is hoped that these findings will help in the development of a synthetic viral vector, which may allow targeted gene therapy in patients with neurological diseases.
Retrograde Cellular Transport of Herpes Simplex Virus
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Description
Herpes simplex virus type 1 (HSV-1) establishes life-long latent infection in sensory neurones, making it an ideal gene therapy vector to target neuronal cells. To study retrograde transport of HSV-1, capsid and tegument proteins were tested for interactions with the motor cytoplasmic dynein, using a yeast two-hybrid system and in vitro pull down assays. A strong interaction was demonstrated between the HSV-1 outer capsid protein VP26 (UL35) and the homologous 14 kDa dynein light chains RP3 and Tctex1. The functional importance of VP26 for retrograde transport was confirmed by microinjecting recombinant HSV-1 capsids into living cells. Capsids containing VP26 moved closer to the cell nucleus, while capsids without VP26 remained in a random distribution. Our results suggest that the HSV-1 outer capsid protein VP26 mediates binding of incoming capsids to the retrograde motor cytoplasmic dynein during cellular infection, through interactions with dynein light chains. It is hoped that these findings will help in the development of a synthetic viral vector, which may allow targeted gene therapy in patients with neurological diseases.
Herpes Simplex Virus: Methods and Protocols provides a wide collection of protocols employed in various levels of herpes virus research, including basic protocols on growing viruses in cell culture...
The herpes group consists of viruses which have been placed together on the basis of a number of distinguishing features that they share in common (ANDRE WES, 1962). All these viruses are relatively...
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