title: Interaction and interdependent packaging of tegument protein UL11 and glycoprotein e of herpes simplex virus.
abstract: The UL11 tegument protein of herpes simplex virus plays a critical role in the secondary envelopment; however, the mechanistic details remain elusive. Here, we report a new function of UL11 in the budding process in which it directs efficient acquisition of glycoprotein E (gE) via a direct interaction. In vitro binding assays showed that the interaction required only the first 28, membrane-proximal residues of the cytoplasmic tail of gE, and the C-terminal 26 residues of UL11. A second, weaker binding site was also found in the N-terminal half of UL11. The significance of the gE-UL11 interaction was subsequently investigated with viral deletion mutants. In the absence of the gE tail, virion packaging of UL11, but not other tegument proteins such as VP22 and VP16, was reduced by at least 80%. Reciprocally, wild-type gE packaging was also drastically reduced by about 87% in the absence of UL11, and this defect could be rescued in trans by expressing U(L)11 at the U(L)35 locus. Surprisingly, a mutant that lacks the C-terminal gE-binding site of UL11 packaged nearly normal amounts of gE despite its strong interaction with the gE tail in vitro, indicating that the interaction with the UL11 N terminus may be important. Mutagenesis studies of the UL11 N terminus revealed that the association of UL11 with membrane was not required for this function. In contrast, the UL11 acidic cluster motif was found to be critical for gE packaging and was not replaceable with foreign acidic clusters. Together, these results highlight an important role of UL11 in the acquisition of glycoprotein-enriched lipid bilayers, and the findings may also have important implications for the role of UL11 in gE-mediated cell-to-cell spread.
authors: Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine, 500 University Drive, P.O. Box 850, Hershey, PA 17033, USA.
authors: Jun
authors: J
authors: Han
authors: Pooja
authors: P
authors: Chadha
authors: David G
authors: DG
authors: Meckes
authors: Nicholas L
authors: NL
authors: Baird
authors: John W
authors: JW
authors: Wills
mesh_terms: D000818:Animals; D002522:Chlorocebus aethiops; D017353:Gene Deletion; D018259:Herpesvirus 1, Human / Q000502:physiology*; D011485:Protein Binding; D025941:Protein Interaction Mapping; D014709:Vero Cells; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism*; D015678:Viral Structural Proteins / Q000235:genetics / Q000378:metabolism*; D019065:Virus Assembly
chemical_list: C073414:UL11 protein, herpesviridae; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins; C044768:glycoprotein E, herpes simplex virus type 1