DNA methyltransferase DNMT3A associates with viral proteins and impacts HSV-1 infection.

TitleDNA methyltransferase DNMT3A associates with viral proteins and impacts HSV-1 infection.
Publication TypeJournal Article
Year of Publication2015
AuthorsRowles, DL, Tsai, Y-C, Greco, TM, Lin, AE, Li, M, Yeh, J, Cristea, IM
JournalProteomics
Volume15
Issue12
Pagination1968-82
Date Published2015 06 01
ISSN1615-9861
KeywordsAnimals, Capsid Proteins, Cells, Cultured, Cercopithecus aethiops, DNA (Cytosine-5-)-Methyltransferase, Fibroblasts, Herpes Simplex, Herpesvirus 1, Human, Host-Pathogen Interactions, Humans, Immunoblotting, Immunoprecipitation, Phosphorylation, Proteome, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Vero Cells, Viral Proteins, Virus Replication
Abstract

Viral infections can alter the cellular epigenetic landscape, through modulation of either DNA methylation profiles or chromatin remodeling enzymes and histone modifications. These changes can act to promote viral replication or host defense. Herpes simplex virus type 1 (HSV-1) is a prominent human pathogen, which relies on interactions with host factors for efficient replication and spread. Nevertheless, the knowledge regarding its modulation of epigenetic factors remains limited. Here, we used fluorescently-labeled viruses in conjunction with immunoaffinity purification and MS to study virus-virus and virus-host protein interactions during HSV-1 infection in primary human fibroblasts. We identified interactions among viral capsid and tegument proteins, detecting phosphorylation of the capsid protein VP26 at sites within its UL37-binding domain, and an acetylation within the major capsid protein VP5. Interestingly, we found a nuclear association between viral capsid proteins and the de novo DNA methyltransferase DNA (cytosine-5)-methyltransferase 3A (DNMT3A), which we confirmed by reciprocal isolations and microscopy. We show that drug-induced inhibition of DNA methyltransferase activity, as well as siRNA- and shRNA-mediated DNMT3A knockdowns trigger reductions in virus titers. Altogether, our results highlight a functional association of viral proteins with the mammalian DNA methyltransferase machinery, pointing to DNMT3A as a host factor required for effective HSV-1 infection.

DOI10.1002/pmic.201500035
Alternate JournalProteomics
PubMed ID25758154
PubMed Central IDPMC4550099
Grant ListDP1 DA026192 / DA / NIDA NIH HHS / United States
R21HD073044 / HD / NICHD NIH HHS / United States
DP1DA026192 / DA / NIDA NIH HHS / United States
R21 AI102187 / AI / NIAID NIH HHS / United States
R01 HL127640 / HL / NHLBI NIH HHS / United States
R21 HD073044 / HD / NICHD NIH HHS / United States
R21AI102187 / AI / NIAID NIH HHS / United States
R01HL127640 / HL / NHLBI NIH HHS / United States