A surface groove essential for viral Bcl-2 function during chronic infection in vivo

Joy Loh, Qiulong Huang, Andrew M. Petros, David Nettesheim, Linda F. Van Dyk, Lucia Labrada, Samuel H. Speck, Beth Levine, Edward T. Olejniczak, Herbert W. Virgin IV

Research output: Contribution to journalArticle

52 Scopus citations

Abstract

Antiapoptotic Bcl-2 family proteins inhibit apoptosis in cultured cells by binding BH3 domains of proapoptotic Bcl-2 family members via a hydrophobic BH3 binding groove on the protein surface. We investigated the physiological importance of the BH3 binding groove of an antiapoptotic Bcl-2 protein in mammals in vivo by analyzing a viral Bcl-2 family protein. We show that the γ-herpesvirus 68 (γHV68) Bcl-2 family protein (γHV68 v-Bcl-2), which is known to inhibit apoptosis in cultured cells, inhibits both apoptosis in primary lymphocytes and Bax toxicity in yeast. Nuclear magnetic resonance determination of the γHV68 v-Bcl-2 structure revealed a BH3 binding groove that binds BH3 domain peptides from proapoptotic Bcl-2 family members Bax and Bak via a molecular mechanism shared with host Bcl-2 family proteins, involving a conserved arginine in the BH3 peptide binding groove. Mutations of this conserved arginine and two adjacent amino acids to alanine (SGR to AAA) within the BH3 binding groove resulted in a properly folded protein that lacked the capacity of the wild-type γHV68 v-Bcl-2 to bind Bax BH3 peptide and to block Bax toxicity in yeast. We tested the physiological importance of this v-Bcl-2 domain during viral infection by engineering viral mutants encoding a v-Bcl-2 containing the SGR to AAA mutation. This mutation resulted in a virus defective for both efficient reactivation of γHV68 from latency and efficient persistent γHV68 replication. These studies demonstrate an essential functional role for amino acids in the BH3 peptide binding groove of a viral Bcl-2 family member during chronic infection.

Original languageEnglish (US)
Article numbere10
Pages (from-to)80-91
Number of pages12
JournalPLoS pathogens
Volume1
Issue number1
DOIs
StatePublished - 2005

ASJC Scopus subject areas

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

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    Loh, J., Huang, Q., Petros, A. M., Nettesheim, D., Van Dyk, L. F., Labrada, L., Speck, S. H., Levine, B., Olejniczak, E. T., & Virgin IV, H. W. (2005). A surface groove essential for viral Bcl-2 function during chronic infection in vivo. PLoS pathogens, 1(1), 80-91. [e10]. https://doi.org/10.1371/journal.ppat.0010010