title: Identification of functional domains within the essential large tegument protein pUL36 of pseudorabies virus.
abstract: Proteins of the capsid proximal tegument are involved in the transport of incoming capsids to the nucleus and secondary envelopment after nuclear egress. Homologs of the essential large capsid proximal tegument protein pUL36 are conserved within the Herpesviridae. They interact with another tegument component, pUL37, and contain a deubiquitinating activity in their N termini which, however, is not essential for virus replication. Whereas an internal deletion of 709 amino acids (aa) within the C-terminal half of the alphaherpesvirus pseudorabies virus (PrV) pUL36 does not impair its function (S. Böttcher, B. G. Klupp, H. Granzow, W. Fuchs, K. Michael, and T. C. Mettenleiter, J. Virol. 80:9910-9915, 2006), deletion of the very C terminus does (J. Lee, G. Luxton, and G. A. Smith, J. Virol. 80:12086-12094, 2006). For further characterization we deleted several predicted functional and structural motifs within PrV pUL36 and analyzed the resulting phenotypes in cell culture and a mouse infection model. Extension of the internal deletion to encompass aa 2087 to 2981 exerted only minor effects on virus replication but resulted in prolonged mean survival times of infected mice. Any additional extension did not yield viable virus. Deletion of an N-terminal region containing the deubiquitinating activity (aa 22 to 248) only slightly impaired viral replication in cell culture but slowed neuroinvasion in our mouse model, whereas a strong impairment of viral replication was observed after simultaneous removal of both nonessential domains. Absence of a region containing two predicted leucine zipper motifs (aa 748 to 991) also strongly impaired virus replication and spread. Thus, we identify several domains within the PrV UL36 protein, which, though not essential, are nevertheless important for virus replication.
authors.affiliation: Institute of Molecular Biology, Friedrich-Loeffler-Institut, Südufer 10, 17493 Greifswald-Insel Riems, Germany.
authors.forename: Sindy
authors.initials: S
authors.lastname: Böttcher
authors.forename: Harald
authors.initials: H
authors.lastname: Granzow
authors.forename: Christina
authors.initials: C
authors.lastname: Maresch
authors.forename: Britta
authors.initials: B
authors.lastname: Möhl
authors.forename: Barbara G
authors.initials: BG
authors.lastname: Klupp
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Mettenleiter
mesh_terms: D000818:Animals; D002460:Cell Line; D019295:Computational Biology; D011558:Herpesvirus 1, Suid / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity / Q000648:ultrastructure; D007668:Kidney / Q000166:cytology / Q000821:virology; D051379:Mice; D008854:Microscopy, Electron; D009154:Mutation; D011557:Pseudorabies / Q000401:mortality / Q000821:virology; D011817:Rabbits; D013329:Structure-Activity Relationship; D010948:Viral Plaque Assay; D015678:Viral Structural Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D015678:Viral Structural Proteins