Binding Partners for the UL11 Tegument Protein of Herpes Simplex Virus Type 1

Joshua S. Loomis, Richard J. Courtney, John W. Wills

Research output: Contribution to journalArticle

83 Citations (Scopus)

Abstract

The product of the UL11 gene of herpes simplex virus type 1 (HSV-1) is a 96-amino-acid tegument protein that accumulates on the cytoplasmic face of internal membranes. Although it is thought to be important for nucleocapsid envelopment and egress, the actual function of this protein is unknown. Previous studies focused on the characterization of sequence elements within the UL11 protein that function in membrane binding and trafficking to the Golgi apparatus. Binding was found to be mediated by two fatty acyl groups (myristate and palmitate), while an acidic cluster and a dileucine motif were identified as being important for the recycling of UL11 from the plasma membrane to the Golgi apparatus. The goal of the experiments described here was to identify and characterize binding partners (viral or cellular) of UL11. Using both immunoprecipitation and glutathione S-transferase (GST) pull-down assays, we identified a 40-kDa protein that specifically associates with UL11 from infected Vero cells. Mutational analyses revealed that the acidic cluster and the dileucine motif are required for this association, whereas the entire second half of UL11 is not. In addition, UL11 homologs from pseudorabies and Marek's disease herpesviruses were also found to be capable of binding to the 40-kDa protein from HSV-1-infected cells, suggesting that the interaction is conserved among alphaherpesviruses. Purification and analysis of the 40-kDa protein by mass spectrometry revealed that it is the product of the U L16 gene, a virion protein reported to be involved in nucleocapsid assembly. Cells transfected with a UL16-green fluorescent protein expression vector produced a protein that was of the expected size, could be pulled down with GST-UL11, and accumulated in a Golgi-like compartment only when coexpressed with UL11, indicating that the interaction does not require any other viral products. These data represent the first steps toward elucidating the network of tegument proteins that UL11 links to membranes.

Original languageEnglish (US)
Pages (from-to)11417-11424
Number of pages8
JournalJournal of virology
Volume77
Issue number21
DOIs
StatePublished - Nov 1 2003

Fingerprint

Human herpesvirus 1
Human Herpesvirus 1
Proteins
proteins
Nucleocapsid
nucleocapsid
Golgi Apparatus
Glutathione Transferase
Golgi apparatus
Membranes
glutathione transferase
Pseudorabies
Marek Disease
Vero Cells
Palmitates
Herpesviridae
Aujeszky disease
Myristic Acid
Recycling
Marek disease

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Loomis, Joshua S. ; Courtney, Richard J. ; Wills, John W. / Binding Partners for the UL11 Tegument Protein of Herpes Simplex Virus Type 1. In: Journal of virology. 2003 ; Vol. 77, No. 21. pp. 11417-11424.
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Binding Partners for the UL11 Tegument Protein of Herpes Simplex Virus Type 1. / Loomis, Joshua S.; Courtney, Richard J.; Wills, John W.

In: Journal of virology, Vol. 77, No. 21, 01.11.2003, p. 11417-11424.

Research output: Contribution to journalArticle

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