Abstract
Histone deacetylation plays a pivotal role in regulating human cytomegalovirus gene expression. In this report, we have identified candidate HDAC1-interacting proteins in the context of infection by using a method for rapid immunoisolation of an epitope-tagged protein coupled with mass spectrometry. Putative interactors included multiple human cytomegalovirus-coded proteins. In particular, the interaction of pUL38 and pUL29/28 with HDAC1 was confirmed by reciprocal immunoprecipitations. HDAC1 is present in numerous protein complexes, including the HDAC1-containing nucleosome remodeling and deacetylase protein complex, NuRD. pUL38 and pUL29/28 associated with the MTA2 component of NuRD, and shRNA-mediated knockdown of the RBBP4 and CHD4 constituents of NuRD inhibited HCMV immediate-early RNA and viral DNA accumulation; together this argues that multiple components of the NuRD complex are needed for efficient HCMV replication. Consistent with a positive acting role for the NuRD elements during viral replication, the growth of pUL29/28- or pUL38-deficient viruses could not be rescued by treating infected cells with the deacetylase inhibitor, trichostatin A. Transient expression of pUL29/28 enhanced activity of the HCMV major immediate-early promoter in a reporter assay, regardless of pUL38 expression. Importantly, induction of the major immediate-early reporter activity by pUL29/28 required functional NuRD components, consistent with the inhibition of immediate-early RNA accumulation within infected cells after knockdown of RBBP4 and CHD4. We propose that pUL29/28 modifies the NuRD complex to stimulate the accumulation of immediate-early RNAs.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Autoantigens / genetics
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Autoantigens / metabolism
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Blotting, Western
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Cells, Cultured
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Chromatin Immunoprecipitation
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Cytomegalovirus / pathogenicity*
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Cytomegalovirus Infections / genetics*
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Cytomegalovirus Infections / metabolism
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Cytomegalovirus Infections / virology
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Fibroblasts / cytology
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Fibroblasts / drug effects
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Fibroblasts / metabolism
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Fluorescent Antibody Technique
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Histone Deacetylase 1 / antagonists & inhibitors
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Histone Deacetylase 1 / genetics
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Histone Deacetylase 1 / metabolism
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Histone Deacetylase Inhibitors / pharmacology
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Histone Deacetylases / genetics
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Histone Deacetylases / metabolism
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Humans
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Hydroxamic Acids / pharmacology
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Immediate-Early Proteins / genetics*
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Immunoprecipitation
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Luciferases / metabolism
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Mi-2 Nucleosome Remodeling and Deacetylase Complex / antagonists & inhibitors
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Mi-2 Nucleosome Remodeling and Deacetylase Complex / genetics
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Mi-2 Nucleosome Remodeling and Deacetylase Complex / metabolism*
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Promoter Regions, Genetic
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RNA, Messenger / genetics
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RNA, Viral / genetics*
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Repressor Proteins / genetics
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Repressor Proteins / metabolism
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Retinoblastoma-Binding Protein 4 / genetics
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Retinoblastoma-Binding Protein 4 / metabolism
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Reverse Transcriptase Polymerase Chain Reaction
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Viral Proteins / genetics
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Viral Proteins / metabolism*
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Virus Replication
Substances
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Autoantigens
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CHD4 protein, human
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Histone Deacetylase Inhibitors
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Hydroxamic Acids
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Immediate-Early Proteins
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RNA, Messenger
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RNA, Viral
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Repressor Proteins
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Retinoblastoma-Binding Protein 4
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Viral Proteins
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trichostatin A
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Luciferases
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MTA2 protein, human
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HDAC1 protein, human
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Histone Deacetylase 1
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Histone Deacetylases
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Mi-2 Nucleosome Remodeling and Deacetylase Complex