A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus

Anna K. Großkopf, Armin Ensser, Frank Neipel, Doris Jungnickl, Sarah Schlagowski, Ronald Charles Desrosiers, Alexander S. Hahn

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Kaposi’s sarcoma-associated herpesvirus (KSHV) is a human oncogenic virus associated with Kaposi’s sarcoma and two B-cell malignancies. The rhesus monkey rhadinovirus (RRV) is a virus of nonhuman primates that is closely related to KSHV. Eph family receptor tyrosine kinases (Ephs) are cellular receptors for the gH/gL glycoprotein complexes of both KSHV and RRV. Through sequence analysis and mutational screens, we identified conserved residues in the N-terminal domain of KSHV and RRV glycoprotein H that are critical for Eph-binding in vitro. Homology-based structural predictions of the KSHV and RRV gH/gL complexes based on the Epstein-Barr-Virus gH/gL crystal structure located these amino acids in a beta-hairpin on gH, which is likely stabilized by gL and is optimally positioned for protein-protein interactions. Guided by these predictions, we generated recombinant RRV and KSHV strains mutated in the conserved motif as well as an RRV gL null mutant. Inhibition experiments using these mutants confirmed that disruption of the identified Eph-interaction motif or of gL expression resulted in complete detargeting from Ephs. However, all mutants were infectious on all cell types tested, exhibiting normal attachment but a reduction in infectivity of up to one log order of magnitude. While Eph-binding-negative RRV mutants were replication-competent on fibroblasts, their infectivity was comparatively more reduced on endothelial cells with a substantial subpopulation of endothelial cells remaining resistant to infection. Together, this provides evidence for a cell type-specific use of Ephs by RRV. Furthermore, our results demonstrate that gL is dispensable for infection by RRV. Its deletion caused a reduction in infectivity similar to that observed after mutation of Eph-binding residues in gH. Our findings would be compatible with an ability of KSHV and RRV to use other, less efficient entry mediators in lieu of Ephs, although these host factors may not be uniformly expressed by all cells.

Original languageEnglish (US)
Article numbere1006912
JournalPLoS Pathogens
Volume14
Issue number2
DOIs
StatePublished - Feb 1 2018

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Rhadinovirus
Eph Family Receptors
Human Herpesvirus 8
Macaca mulatta
Glycoproteins
Endothelial Cells
Oncogenic Viruses
Kaposi's Sarcoma
Infection
Human Herpesvirus 4
Primates
Sequence Analysis
Proteins
B-Lymphocytes

ASJC Scopus subject areas

  • Parasitology
  • Microbiology
  • Immunology
  • Molecular Biology
  • Genetics
  • Virology

Cite this

A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus. / Großkopf, Anna K.; Ensser, Armin; Neipel, Frank; Jungnickl, Doris; Schlagowski, Sarah; Desrosiers, Ronald Charles; Hahn, Alexander S.

In: PLoS Pathogens, Vol. 14, No. 2, e1006912, 01.02.2018.

Research output: Contribution to journalArticle

Großkopf, Anna K. ; Ensser, Armin ; Neipel, Frank ; Jungnickl, Doris ; Schlagowski, Sarah ; Desrosiers, Ronald Charles ; Hahn, Alexander S. / A conserved Eph family receptor-binding motif on the gH/gL complex of Kaposi’s sarcoma-associated herpesvirus and rhesus monkey rhadinovirus. In: PLoS Pathogens. 2018 ; Vol. 14, No. 2.
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AU - Neipel, Frank

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AU - Hahn, Alexander S.

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