title: Properties of the protein encoded by the UL32 open reading frame of herpes simplex virus 1.
abstract: The functions previously assigned to the essential herpes simplex virus 1 UL32 protein were in cleavage and/or packaging of viral DNA and in maturation and/or translocation of viral glycoproteins to the plasma membrane. The amino acid sequence predicts N-linked glycosylation sites and sequences conserved in aspartyl proteases and in zinc-binding proteins. We report the following. (i) The 596-amino-acid UL32 protein accumulated predominantly in the cytoplasm of infected cells but was not metabolically labeled with glucosamine and did not band with membranes containing a known glycoprotein in flotation sucrose density gradients. The UL32 protein does not, therefore, have the properties of an intrinsic membrane protein. (ii) Experiments designed to demonstrate aspartyl protease activity in a phage display system failed to reveal proteolytic activity. Moreover, substitution of Asp-110 with Gly in the sequence Asp-Thr-Gly, the hallmark of aspartyl proteases, had no effect on viral replication in Vero and SK-N-SH cell lines or in human foreskin fibroblasts. Therefore, if the UL32 protein functions as a protease, this function is not required in cells in culture. (iii) Both the native UL32 protein and a histidine-tagged UL32 protein made in recombinant baculovirus-infected insect cells bound zinc. The consensus sequence is conserved in the UL32 homologs from varicella-zoster virus and equine herpesvirus 1. UL32 protein is therefore a cysteine-rich, zinc-binding essential cytoplasmic protein whose function is not yet clear.
authors.affiliation: The Marjorie B. Kovler Viral Oncology Laboratories, The University of Chicago, Illinois 60637, USA.
authors.forename: Y E
authors.initials: YE
authors.lastname: Chang
authors.forename: A P
authors.initials: AP
authors.lastname: Poon
authors.forename: B
authors.initials: B
authors.lastname: Roizman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D006224:Cricetinae; D003593:Cytoplasm / Q000737:chemistry; D010450:Endopeptidases / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014764:Viral Proteins / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D014779:Virus Replication; D015032:Zinc / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D007106:Immune Sera; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C069185:DNA cleavage and packaging proteins, Herpesvirus; D010450:Endopeptidases; D015032:Zinc