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Human cytomegalovirus pUL97 regulates the viral major immediate early promoter by phosphorylation-mediated disruption of histone deacetylase 1 binding. J Virol 2013 Jul;87(13):7393-408

Date

04/26/2013

Pubmed ID

23616659

Pubmed Central ID

PMC3700266

DOI

10.1128/JVI.02825-12

Scopus ID

2-s2.0-84880367640   23 Citations

Abstract

Human cytomegalovirus (HCMV) is a common agent of congenital infection and causes severe disease in immunocompromised patients. Current approved therapies focus on inhibiting viral DNA replication. The HCMV kinase pUL97 contributes to multiple stages of viral infection including DNA replication, controlling the cell cycle, and virion maturation. Our studies demonstrate that pUL97 also functions by influencing immediate early (IE) gene expression during the initial stages of infection. Inhibition of kinase activity using the antiviral compound maribavir or deletion of the UL97 gene resulted in decreased expression of viral immediate early genes during infection. Expression of pUL97 was sufficient to transactivate IE1 gene expression from the viral genome, which was dependent on viral kinase activity. We observed that pUL97 associates with histone deacetylase 1 (HDAC1). HDAC1 is a transcriptional corepressor that acts to silence expression of viral genes. We observed that inhibition or deletion of pUL97 kinase resulted in increased HDAC1 and decreased histone H3 lysine 9 acetylation associating with the viral major immediate early (MIE) promoter. IE expression during pUL97 inhibition or deletion was rescued following inhibition of deacetylase activity. HDAC1 associates with chromatin by protein-protein interactions. Expression of active but not inactive pUL97 kinase decreased HDAC1 interaction with the transcriptional repressor protein DAXX. Finally, using mass spectrometry, we found that HDAC1 is uniquely phosphorylated upon expression of pUL97. Our results support the conclusion that HCMV pUL97 kinase regulates viral immediate early gene expression by phosphorylation-mediated disruption of HDAC1 binding to the MIE promoter.

Author List

Bigley TM, Reitsma JM, Mirza SP, Terhune SS

Author

Scott Terhune PhD Professor in the Microbiology and Immunology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Adaptor Proteins, Signal Transducing
Benzimidazoles
Blotting, Western
Chromatin Immunoprecipitation
Co-Repressor Proteins
Cytomegalovirus
DNA Primers
Fluorescent Antibody Technique
Gene Deletion
Gene Expression Regulation, Viral
Histone Deacetylase 1
Humans
Immediate-Early Proteins
Immunoprecipitation
Mass Spectrometry
Molecular Chaperones
Mutagenesis, Site-Directed
Nuclear Proteins
Phosphorylation
Protein Kinases
Ribonucleosides