Human cytomegalovirus UL29/28 protein interacts with components of the NuRD complex which promote accumulation of immediate-early RNA. PLoS Pathog 2010 Jun 24;6(6):e1000965
Date
06/30/2010Pubmed ID
20585571Pubmed Central ID
PMC2891856DOI
10.1371/journal.ppat.1000965Scopus ID
2-s2.0-77954671476 (requires institutional sign-in at Scopus site) 65 CitationsAbstract
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.
Author List
Terhune SS, Moorman NJ, Cristea IM, Savaryn JP, Cuevas-Bennett C, Rout MP, Chait BT, Shenk TAuthor
Scott Terhune PhD Professor in the Microbiology and Immunology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AutoantigensBlotting, Western
Cells, Cultured
Chromatin Immunoprecipitation
Cytomegalovirus
Cytomegalovirus Infections
Fibroblasts
Fluorescent Antibody Technique
Histone Deacetylase 1
Histone Deacetylase Inhibitors
Histone Deacetylases
Humans
Hydroxamic Acids
Immediate-Early Proteins
Immunoprecipitation
Luciferases
Mi-2 Nucleosome Remodeling and Deacetylase Complex
Promoter Regions, Genetic
RNA, Messenger
RNA, Viral
Repressor Proteins
Retinoblastoma-Binding Protein 4
Reverse Transcriptase Polymerase Chain Reaction
Viral Proteins
Virus Replication