Differential relocation and stability of PML-body components during productive human cytomegalovirus infection: detailed characterization by live-cell imaging

Eur J Cell Biol. 2010 Oct;89(10):757-68. doi: 10.1016/j.ejcb.2010.05.006.

Abstract

In controlling the switch from latency to lytic infection, the immediate early (IE) genes lie at the core of herpesvirus pathogenesis. To image the 72kDa human cytomegalovirus (HCMV) major IE protein (IE1-72K), a recombinant virus encoding IE1 fused with EGFP was constructed. Using this construct, the IE1-EGFP fusion was detected at ND10 (PML-bodies) within 2h post infection (p.i.) and the complete disruption of ND10 imaged through to 6h p.i. HCMV genomes and IE2-86K protein could be detected adjacent to the slowly degrading IE1-72K/ND10 foci. IE1-72K associates with metaphase chromatin, recruiting both PML and STAT2. hDaxx, STAT1 and IE2-86K did not re-locate to metaphase chromatin; the fate of hDaxx is particularly important as this protein contributes to an intrinsic barrier to HCMV infection. While IE1-72K participates in a complex with chromatin, PML, STAT2 and Sp100, IE1-72K releases hDaxx from ND10 yet does not appear to remain associated with it.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens, Nuclear / genetics
  • Antigens, Nuclear / metabolism
  • Autoantigens / genetics
  • Autoantigens / metabolism
  • Chromatin / chemistry
  • Chromatin / genetics
  • Chromatin / metabolism
  • Cytomegalovirus / genetics
  • Cytomegalovirus / metabolism*
  • Cytomegalovirus Infections / genetics
  • Cytomegalovirus Infections / metabolism
  • HeLa Cells
  • Humans
  • Image Processing, Computer-Assisted
  • Immediate-Early Proteins / chemistry
  • Immediate-Early Proteins / genetics
  • Immediate-Early Proteins / metabolism*
  • Metaphase / genetics
  • Microscopy, Fluorescence
  • Microscopy, Video
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Promyelocytic Leukemia Protein
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • STAT2 Transcription Factor / genetics
  • STAT2 Transcription Factor / metabolism
  • Trans-Activators / genetics
  • Trans-Activators / metabolism*
  • Transcription Factors / chemistry
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • Tumor Suppressor Proteins / chemistry
  • Tumor Suppressor Proteins / genetics
  • Tumor Suppressor Proteins / metabolism*

Substances

  • Antigens, Nuclear
  • Autoantigens
  • CALCOCO2 protein, human
  • Chromatin
  • IE1 protein, cytomegalovirus
  • IE2 protein, Cytomegalovirus
  • Immediate-Early Proteins
  • Nuclear Proteins
  • Promyelocytic Leukemia Protein
  • Recombinant Fusion Proteins
  • STAT2 Transcription Factor
  • STAT2 protein, human
  • Trans-Activators
  • Transcription Factors
  • Tumor Suppressor Proteins
  • SP100 protein, human
  • PML protein, human