title: Characterization of the herpes simplex virus (HSV)-1 tegument protein VP1-2 during infection with the HSV temperature-sensitive mutant tsB7.
abstract: VP1-2, encoded by the UL36 gene of herpes simplex virus (HSV), is a large structural protein, conserved across the family Herpesviridae, that is assembled into the tegument and is essential for virus replication. Current evidence indicates that VP1-2 is a central component in the tegumentation and envelopment processes and that it also possesses important roles in capsid transport and entry. However, any detailed mechanistic understanding of VP1-2 function(s) remains limited. This study characterized the replication of HSV-1 tsB7, a temperature-sensitive mutant restricted at the non-permissive temperature due to a defect in VP1-2 function. A tsB7 virus expressing green fluorescent protein-fused VP16 protein was used to track the accumulation and location of a major tegument protein. After infection at the permissive temperature and shift to the non-permissive temperature, the production of infectious virus ceased. VP1-2 accumulated in altered cytosolic clusters, together with VP16 and other virion proteins. Furthermore, correlating with the results of immunofluorescence, electron microscopy demonstrated abnormal cytosolic capsid clustering and a block in envelopment. As VP1-2 encompasses a ubiquitin-specific protease domain, the occurrence of ubiquitin-conjugated proteins during tsB7 infection was also examined at the non-permissive temperature. A striking overaccumulation was observed of ubiquitin-specific conjugates in cytoplasmic clusters, overlapping and adjacent to the VP1-2 clusters. These results are discussed in relation to the possible functions of VP1-2 in the assembly pathway and the nature of the defect in tsB7.
authors: Marie Curie Research Institute, The Chart, Oxted RH8 0TL, Surrey, UK.
authors: F
authors: F
authors: Abaitua
authors: Division of Virology, Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, UK.
authors: R N
authors: RN
authors: Souto
authors: Division of Virology, Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, UK.
authors: H
authors: H
authors: Browne
authors: Marie Curie Research Institute, The Chart, Oxted RH8 0TL, Surrey, UK.
authors: T
authors: T
authors: Daikoku
authors: Marie Curie Research Institute, The Chart, Oxted RH8 0TL, Surrey, UK.
authors: P
authors: P
authors: O'Hare
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D049452:Green Fluorescent Proteins; D018259:Herpesvirus 1, Human / Q000378:metabolism*; D006801:Humans; D009154:Mutation; D021381:Protein Transport; D011994:Recombinant Proteins; D013696:Temperature; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins