Bromodomain and extraterminal inhibitors block the Epstein-Barr virus lytic cycle at two distinct steps

Keck, Kristin M.; Moquin, S.; He, Amanda; Fernandez, Samantha G.; Somberg, Jessica J.; Liu, Stephanie M.; Martinez, Delsy; Miranda, JJ L.

doi:10.1074/jbc.m116.751644

Cited by 30 publications

(33 citation statements)

References 43 publications

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“…Although we did not monitor viral DNA replication or other viral transcripts, the transient nature of the assay (2 days infection to permit viral entry followed by 3 days of drug treatment) could suggest that the anti-viral effect is post-entry and my potentially target both the initial but limited unlicensed viral DNA replication as well as early viral mRNA expression that occurs when virus particles infect basal keratinocytes (McBride and Jang, 2013). Interestingly it has been shown that a BET inhibitor blocked replication of EBV both by blocking early viral gene expression and lytic genome replication (Keck et al, 2017). …”

Section: Discussionmentioning

confidence: 99%

“…The role BET proteins play in the life cycle of viruses has been explored for other DNA viruses and retroviruses and BET inhibitors show contrasting effects depending on the virus, for example BET inhibitors reactivated HIV in models of latency and blocked EBV early gene expression and lytic genome replication (Banerjee et al, 2012; Keck et al, 2017; Palermo et al, 2011; Weidner-Glunde et al, 2010; Zhu et al, 2012). Since BRD4/PTEFb dependent activation of HPV oncogenes has been described the effect of BET inhibitors on reducing HPV oncogene expression could also be worthwhile investigating (Yan et al, 2010).…”

Section: Discussionmentioning

confidence: 99%

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BET bromodomain inhibitors show anti-papillomavirus activity in vitro and block CRPV wart growth in vivo

et al. 2018

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The DNA papillomaviruses infect squamous epithelium and can cause persistent, benign and sometimes malignant hyperproliferative lesions. Effective antiviral drugs to treat human papillomavirus (HPV) infection are lacking and here we investigate the anti-papillomavirus activity of novel epigenetic targeting drugs, BET bromodomain inhibitors. Bromodomain and Extra-Terminal domain (BET) proteins are host proteins which regulate gene transcription, they bind acetylated lysine residues in histones and non-histone proteins via bromodomains, functioning as scaffold proteins in the formation of transcriptional complexes at gene regulatory regions. The BET protein BRD4 has been shown to be involved in the papillomavirus life cycle, as a co-factor for viral E2 and also mediating viral partitioning in some virus types. We set out to study the activity of small molecule BET bromodomain inhibitors in models of papillomavirus infection. Several BET inhibitors reduced HPV11 E1ˆE4 mRNA expression in vitro and topical therapeutic administration of an exemplar compound I-BET762, abrogated CRPV cutaneous wart growth in rabbits, demonstrating translation of anti-viral effects to efficacy in vivo. Additionally I-BET762 markedly reduced viability of HPV16 infected W12 cells compared to non-infected C33A cells. The molecular mechanism for the cytotoxicity to W12 cells is unknown but may be through blocking viral-dependent cell-survival factors. We conclude that these effects, across multiple papillomavirus types and in vivo, highlight the potential to target BET bromodomains to treat HPV infection.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

BET bromodomain inhibitors show anti-papillomavirus activity in vitro and block CRPV wart growth in vivo

et al. 2018

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“…DNA deep sequencing of acyclovir---and doxycycline---treated LNGFR---cells, which represent the latent population, estimate a viral genome copy number of 20. Acyclovir indeed functions as intended; total EBV DNA content increased by only 2.1±0.4---fold, much of which comes from abortive replication attempts near the lytic origins (30). Comparison of EBV episomes in latent and lytic cells revealed that the chromosome association preferences are lost during reactivation ( Fig 6A and 6B).…”

Section: Introductionmentioning

confidence: 87%

“…Genomic DNA was purified by silica---based membrane affinity with a DNeasy Blood & Tissue Kit (Qiagen). Libraries were constructed and the percentage of EBV reads from total measured (30). Viral genome copy number was estimated based on observed ratios between the number of reads mapped to the EBV and human genome sequences compared to the expected value, which is calculated as the ratio between the viral episome length and the summed length of all human chromosomes.…”

Section: Analysis Of Viral---human Contact Regionsmentioning

confidence: 99%

The Epstein-Barr virus episome maneuvers between nuclear chromatin compartments during reactivation

Moquin

Thomas

Whalen

et al. 2017

Preprint

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The human genome is structurally organized in three---dimensional space to facilitate functional partitioning of transcription. We learned that the latent episome of the human Epstein---Barr virus (EBV) preferentially associates with gene---poor chromosomes and avoids gene---rich chromosomes. Kaposi's sarcoma---associated herpesvirus behaves similarly, but human papillomavirus does not. Contacts localize on the EBV side to OriP, the latent origin of replication. This genetic element, and the EBNA1 protein that binds there, are sufficient to reconstitute chromosome association preferences of the entire episome. Contacts localize on the human side to gene---poor regions of chromatin distant from transcription start sites. Upon reactivation from latency, however, the episome moves away from repressive heterochromatin and toward active euchromatin. Our work adds three---dimensional relocalization to the molecular events that occur during reactivation. Involvement of a myriad of interchromosomal associations also suggests a role for this type of long---range association in gene regulation. IMPORTANCEThe human genome is structurally organized in three---dimensional space, and this structure functionally affects transcriptional activity. We set out to investigate whether a double stranded DNA virus, Epstein---Barr virus (EBV), uses similar mechanisms as the human genome to regulate transcription. We found that the EBV genome associates with repressive compartments of the nucleus during latency and active compartments during reactivation. This study is advances our knowledge of the EBV life cycle, adding three---dimensional re---localization as a novel component to the molecular events that occur during reactivation. Furthermore, the data adds to our understanding of nuclear compartments, showing that disperse interchromosomal interactions may be important for regulating transcription.

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“…One potential technique is generating EBV-positive cells that harbor a truncated version of the human low-affinity nerve growth factor receptor (LNGFR) molecule as a cell surface marker driven by either an early or a late promoter. For example, EBV-positive cells, in which LNGFR expression is under the control of a tetracycline-dependent promoter that also drives the expression of BZLF1, were selected using anti-LNGFR magnetically activated cell sorting (MACS) beads (85,86). Similarly, the rat CD-2 protein driven by an EBV early promoter allowed the selection of cells-by affinity purification-in which EBV was entering its lytic cycle (87).…”

mentioning

confidence: 99%

An Epigenetic Journey: Epstein-Barr Virus Transcribes Chromatinized and Subsequently Unchromatinized Templates during Its Lytic Cycle

Chakravorty

Sugden

Johannsen

2019

J Virol

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The Epstein-Barr virus (EBV) lytic phase, like those of all herpesviruses, proceeds via an orderly cascade that integrates DNA replication and gene expression. EBV early genes are expressed independently of viral DNA amplification, and several early gene products facilitate DNA amplification. On the other hand, EBV late genes are defined by their dependence on viral DNA replication for expression. Recently, a set of orthologous genes found in beta-and gammaherpesviruses have been determined to encode a viral preinitiation complex (vPIC) that mediates late gene expression. The EBV vPIC requires an origin of lytic replication in cis, implying that the vPIC mediates transcription from newly replicated DNA. In agreement with this implication, EBV late gene mRNAs localize to replication factories. Notably, these factories exclude canonical histones. In this review, we compare and contrast the mechanisms and epigenetics of EBV early and late gene expression. We summarize recent findings, propose a model explaining the dependence of EBV late gene expression on lytic DNA amplification, and suggest some directions for future study.with 5-ethynyl uridine (EU) in the early and late lytic phases, followed by enrichment for EU-labeled RNAs and RNA-seq to assay for viral and cellular genes that are being actively transcribed at different times during the lytic cycle. Similar experiments using ribosome profiling may yield valuable information about which mRNAs are being actively translated during the lytic phase. For example, Bencun et al. (88) used ribosome profiling on EBV-positive cells to show highly variable translation of EBV transcripts. However, only about 4 to 6% of the cells used in the assay were in the lytic phase. TheFIG 3 A model for EBV late gene expression. (A) Early during the lytic cycle, EBV genomes are unreplicated and still chromatinized. The immediate early proteins BZLF1 and BRLF1 can bind to the promoters of early genes (which are usually methylated) and, along with cellular TATA box-binding protein (TBP), recruit RNA polymerase II to transcribe early genes. (B) Later in the lytic phase, EBV genomes are amplified in replication factories, while cellular chromatin moves to the periphery of the nucleus. The amplified viral DNA is unmethylated and unchromatinized, which allows the viral preinitiation complex (vPIC), including the viral TBP BcRF1, to recruit RNA Pol II to late promoters and transcribe late genes.

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Bromodomain and extraterminal inhibitors block the Epstein-Barr virus lytic cycle at two distinct steps

Cited by 30 publications

References 43 publications

BET bromodomain inhibitors show anti-papillomavirus activity in vitro and block CRPV wart growth in vivo

BET bromodomain inhibitors show anti-papillomavirus activity in vitro and block CRPV wart growth in vivo

The Epstein-Barr virus episome maneuvers between nuclear chromatin compartments during reactivation

An Epigenetic Journey: Epstein-Barr Virus Transcribes Chromatinized and Subsequently Unchromatinized Templates during Its Lytic Cycle

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