Identification of human cytomegalovirus genes important for biogenesis of the cytoplasmic virion assembly complex

Subhendu Das, Daniel A. Ortiz, Stephen J. Gurczynski, Fatin Khan, Philip E. Pellett

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Human cytomegalovirus (HCMV) has many effects on cells, including remodeling the cytoplasm to form the cytoplasmic virion assembly complex (cVAC), the site of final virion assembly. Viral tegument, envelope, and some nonstructural proteins localize to the cVAC, and cytoskeletal filaments radiate from a microtubule organizing center in the cVAC. The endoplasmic reticulum (ER)-to-Golgi intermediate compartment, Golgi apparatus, and trans-Golgi network form a ring that outlines the cVAC. The center of the cVAC ring is occupied by numerous vesicles that share properties with recycling endosomes. In prior studies, we described the three-dimensional structure and the extensive remodeling of the cytoplasm and shifts in organelle identity that occur during development of the cVAC. The objective of this work was to identify HCMV proteins that regulate cVAC biogenesis. Because the cVAC does not form in the absence of viral DNA synthesis, we employed HCMV-infected cells transfected with synthetic small interfering RNAs (siRNAs) that targeted 26 candidate early-late and late protein-coding genes required for efficient virus replication. We identified three HCMV genes (UL48, UL94, and UL103) whose silencing had major effects on cVAC development, including failure to form the Golgi ring and dispersal of markers of early and recycling endosomes. To confirm and extend the siRNA results, we constructed recombinant viruses in which pUL48 and pUL103 are fused with a regulatable protein destabilization domain (dd-FKBP). In the presence of a stabilizing ligand (Shield-1), the cVAC appeared to develop normally. In its absence, cVAC development was abrogated, verifying roles for pUL48 and pUL103 in cVAC biogenesis.

Original languageEnglish (US)
Pages (from-to)9086-9099
Number of pages14
JournalJournal of Virology
Volume88
Issue number16
DOIs
StatePublished - Jan 1 2014
Externally publishedYes

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Human herpesvirus 5
Forensic Anthropology
Cytomegalovirus
virion
Virion
Genes
genes
endosomes
Endosomes
small interfering RNA
biogenesis
Small Interfering RNA
recycling
Cytoplasm
cytoplasm
proteins
Microtubule-Organizing Center
Tacrolimus Binding Proteins
trans-Golgi Network
Proteins

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Identification of human cytomegalovirus genes important for biogenesis of the cytoplasmic virion assembly complex. / Das, Subhendu; Ortiz, Daniel A.; Gurczynski, Stephen J.; Khan, Fatin; Pellett, Philip E.

In: Journal of Virology, Vol. 88, No. 16, 01.01.2014, p. 9086-9099.

Research output: Contribution to journalArticle

Das, Subhendu ; Ortiz, Daniel A. ; Gurczynski, Stephen J. ; Khan, Fatin ; Pellett, Philip E. / Identification of human cytomegalovirus genes important for biogenesis of the cytoplasmic virion assembly complex. In: Journal of Virology. 2014 ; Vol. 88, No. 16. pp. 9086-9099.
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