Characterization of pUL5, an HCMV protein interacting with the cellular protein IQGAP1

Virology. 2020 Jan 15:540:57-65. doi: 10.1016/j.virol.2019.10.018. Epub 2019 Nov 3.

Abstract

Among the Herpesviridae, human cytomegalovirus (HCMV) owns the largest genome and displays a huge coding potential. Here, we characterized the UL5 gene product (pUL5) of the clinical isolate TR strain. The protein was predicted as a 166-amino-acid membrane protein with a theoretical mass of 19 kDa. Recombinant virus expressing pUL5 with a tag allowed the identification of two pUL5 non-glycosylated species of approximately 19 and 9 kDa, expressed with early and late kinetic respectively. Experiments in infection confirmed that the lower molecular weight species was translated from an internal ATG in the UL5 open reading frame. Confocal microscopy analysis showed that pUL5 localized within the assembly compartment, but is not incorporated in the virion, as shown by Western blot on purified viral particles. Finally, pull-down experiments coupled with mass spectrometry analysis identified IQGAP1 as a pUL5 interactor, giving new hints on possible roles of pUL5 during HCMV infection.

Keywords: Coding potential; Human cytomegalovirus; IQGAP1; Internal AUG; Non-structural protein; UL5; UL5 isoforms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Cell Line
  • Cells, Cultured
  • Cytomegalovirus / isolation & purification
  • Cytomegalovirus / physiology*
  • Cytomegalovirus / ultrastructure
  • Cytomegalovirus Infections / metabolism*
  • Cytomegalovirus Infections / virology*
  • Gene Expression Regulation, Viral
  • Host-Pathogen Interactions*
  • Humans
  • Open Reading Frames
  • Protein Binding
  • Protein Transport
  • RNA, Viral
  • Transcription, Genetic
  • Viral Proteins / metabolism*
  • ras GTPase-Activating Proteins / metabolism*

Substances

  • IQ motif containing GTPase activating protein 1
  • RNA, Viral
  • Viral Proteins
  • ras GTPase-Activating Proteins