Human cytomegalovirus interactome analysis identifies degradation hubs, domain associations and viral protein functions

Elife. 2019 Dec 24;8:e49894. doi: 10.7554/eLife.49894.

Abstract

Human cytomegalovirus (HCMV) extensively modulates host cells, downregulating >900 human proteins during viral replication and degrading ≥133 proteins shortly after infection. The mechanism of degradation of most host proteins remains unresolved, and the functions of many viral proteins are incompletely characterised. We performed a mass spectrometry-based interactome analysis of 169 tagged, stably-expressed canonical strain Merlin HCMV proteins, and two non-canonical HCMV proteins, in infected cells. This identified a network of >3400 virus-host and >150 virus-virus protein interactions, providing insights into functions for multiple viral genes. Domain analysis predicted binding of the viral UL25 protein to SH3 domains of NCK Adaptor Protein-1. Viral interacting proteins were identified for 31/133 degraded host targets. Finally, the uncharacterised, non-canonical ORFL147C protein was found to interact with elements of the mRNA splicing machinery, and a mutational study suggested its importance in viral replication. The interactome data will be important for future studies of herpesvirus infection.

Keywords: computational biology; host-pathogen interaction; human; human cytomegalovirus; immune evasion; infectious disease; microbiology; protein-protein interaction; proteomics; systems biology; systems virology; virus.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / genetics*
  • Cytomegalovirus / genetics*
  • Cytomegalovirus / pathogenicity
  • Cytomegalovirus Infections / genetics*
  • Cytomegalovirus Infections / virology
  • Gene Expression Regulation, Viral / genetics
  • Host-Pathogen Interactions / genetics
  • Humans
  • Oncogene Proteins / genetics*
  • Proteomics*
  • RNA Splicing / genetics
  • RNA, Messenger / genetics
  • Viral Proteins / genetics
  • Virus Replication / genetics

Substances

  • Adaptor Proteins, Signal Transducing
  • Nck protein
  • Oncogene Proteins
  • RNA, Messenger
  • Viral Proteins