Journal
JOURNAL OF VIROLOGY
Volume 81, Issue 18, Pages 10137-10150Publisher
AMER SOC MICROBIOLOGY
DOI: 10.1128/JVI.01233-06
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Funding
- NATIONAL CANCER INSTITUTE [R01CA083525, R01CA091791, R01CA094809] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES [R56AI012601, R01AI012601, R01AI029733, R01AI046420] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF DENTAL &CRANIOFACIAL RESEARCH [R01DE014153] Funding Source: NIH RePORTER
- NCI NIH HHS [R01 CA091791, CA 91791, CA 83525, CA 94809, R01 CA094809] Funding Source: Medline
- NIAID NIH HHS [R01 AI029733, R56 AI012601, R01 AI046420, R01 AI012601, AI 12601, AI 29733, AI 46420] Funding Source: Medline
- NIDCR NIH HHS [R01 DE014153, DE 14153] Funding Source: Medline
- NIGMS NIH HHS [T32 GM007185, GM 07185] Funding Source: Medline
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The tegument, a semiordered matrix of proteins overlying the nucleocapsid and underlying the virion envelope, in viruses in the gamma subfamily of Herpesviridae is poorly understood. Murine gammaherpesvirus 68 MHV-68, is a robust model for studying gammaherpesvirus virion structure, assembly, and composition, as MHV-68 efficiently completes the lytic phase and productively infects cultured cells. We have found that MHV-68 ORF52 encodes an abundant tegument protein conserved among gammaherpesviruses. Detergent sensitivity experiments revealed that the MIIV-68 ORF52 protein is more tightly bound to the virion nucleocapsid than the ORF45 tegument protein but could be dissociated from particles that retained the ORF65 small capsomer protein. ORF52, tagged with enhanced green fluorescent protein or FLAG epitope, localized to the cytoplasm. A recombinant MHV-68 bacterial artificial chromosome mutant with a nonsense mutation incorporated into ORF52 exhibited viral DNA replication, expression of late lytic genes, and capsid assembly and packaging at levels near those of the wild type. However, the MHV-68 ORF52-null virus was deficient in the assembly and release of infectious virion particles. Instead, partially tegumented capsids produced by the ORF52-null mutant accumulated in the cytoplasm, containing conserved capsid proteins, the ORF64 and ORF67 tegument proteins, but virtually no OPF45 tegument protein. Thus, ORF52 is essential for the tegumentation and egress of infectious MHV-68 particles in the cytoplasm, suggesting an important conserved function in gammaherpesvirus virion morphogenesis.
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