The assembly of human cytomegalovirus (HCMV) is extraordinarily complex requiring the organization and incorporation of over 100 structural proteins. The final step in assembly of the infectious particle requires acquisition of an envelope by the tegumented DNA containing capsid. In previous studies the site of envelopment has been suggested to be a membranous compartment in the cytoplasm of the infected cell, the assembly compartment (AC). From studies of the trafficking of viral glycoproteins and tegument proteins to AC, we have suggested that the AC represents a virus induced compartment at the intersection of anterograde (biosynthetic) and retrograde (endocytic) trafficking pathways marked by cellular proteins of trans-Golgi network (TGN) and the endocytic recycling compartment (ERC). In this proposal we will utilize a variety of biochemical, imaging, and genetic approaches to investigate the role of HCMV viral envelope glycoprotein trafficking through the endocytic pathway to localization in the AC and subsequently to virus assembly. In the second section of the project we will define components of the outer tegument layer of the virion and define pathways of their localization to the AC. In the final section of the proposal, w will determine requirements for production of enveloped particles, including identifying envelope glycoproteins that are essential for particle envelopment and defining interactions between envelope glycoproteins and outer tegument proteins required for virion envelopment in the cytoplasm of infected cells. These studies will provide fundamental information about HCMV assembly and identify potential targets and pathways for antiviral drug development and approaches for the production of non- replicating, virus like particle based vaccines with enhanced immunogenicity and safety profiles.

Public Health Relevance

Studies outlined in this project will provide new insight into cytoplasmic assemble of human cytomegalovirus (HCMV), the largest and structurally most complex human herpesvirus and an important human pathogen. Utilizing a combination of biochemical, imaging, and genetic approaches, this project will investigate the protein interactions required for envelopment of this virus. Results from these studies will point to new targets and strategies for development of therapeutics to limit disease associated with HCMV investigation and improve current approaches in vaccine development.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
2R01AI035602-16A1
Application #
8787824
Study Section
Special Emphasis Panel (ZRG1)
Program Officer
Beisel, Christopher E
Project Start
Project End
Budget Start
Budget End
Support Year
16
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of Alabama Birmingham
Department
Pediatrics
Type
Schools of Medicine
DUNS #
City
Birmingham
State
AL
Country
United States
Zip Code
35294
Rebmann, G Michael; Grabski, Robert; Sanchez, Veronica et al. (2016) Phosphorylation of Golgi Peripheral Membrane Protein Grasp65 Is an Integral Step in the Formation of the Human Cytomegalovirus Cytoplasmic Assembly Compartment. MBio 7:
Seo, Jun-Young; Jeon, Hyejin; Hong, Sookyung et al. (2016) Distinct functional domains within the acidic cluster of tegument protein pp28 required for trafficking and cytoplasmic envelopment of human cytomegalovirus. J Gen Virol 97:2677-2683
Chiuppesi, Flavia; Wussow, Felix; Johnson, Erica et al. (2015) Vaccine-Derived Neutralizing Antibodies to the Human Cytomegalovirus gH/gL Pentamer Potently Block Primary Cytotrophoblast Infection. J Virol 89:11884-98
Hook, Lauren M; Grey, Finn; Grabski, Robert et al. (2014) Cytomegalovirus miRNAs target secretory pathway genes to facilitate formation of the virion assembly compartment and reduce cytokine secretion. Cell Host Microbe 15:363-73
Hook, Lauren; Hancock, Meaghan; Landais, Igor et al. (2014) Cytomegalovirus microRNAs. Curr Opin Virol 7:40-6
Indran, Sabarish V; Britt, William J (2011) A role for the small GTPase Rab6 in assembly of human cytomegalovirus. J Virol 85:5213-9
Yu, Xuekui; Shah, Sanket; Lee, Manfred et al. (2011) Biochemical and structural characterization of the capsid-bound tegument proteins of human cytomegalovirus. J Struct Biol 174:451-60
Indran, Sabarish V; Ballestas, Mary E; Britt, William J (2010) Bicaudal D1-dependent trafficking of human cytomegalovirus tegument protein pp150 in virus-infected cells. J Virol 84:3162-77
Kropff, Barbara; Koedel, Yvonne; Britt, William et al. (2010) Optimal replication of human cytomegalovirus correlates with endocytosis of glycoprotein gpUL132. J Virol 84:7039-52
Seo, Jun-Young; Britt, William J (2008) Multimerization of tegument protein pp28 within the assembly compartment is required for cytoplasmic envelopment of human cytomegalovirus. J Virol 82:6272-87

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