Cross-species conservation of episome maintenance provides a basis for in vivo investigation of Kaposi's sarcoma herpesvirus LANA

Habison, A. C.; Miranda, Marta Pires de; Beauchemin, Chantal; Tan, Min; Cerqueira, Sofia; Correia, Bruno E.; Ponnusamy, Rajesh; Usherwood, Edward J.; McVey, Colin E.; Simas, J. Pedro; Kaye, Kenneth M.

doi:10.1371/journal.ppat.1006555

Cited by 20 publications

(48 citation statements)

References 73 publications

(87 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Under antibiotic selection, kLANA as well as mLANA-expressing cells (but not the parental ML12 cells) yielded hygromycin-resistant cultures, as was expected based on the previously reported ability of mLANA to support maintenance of episomes harboring KSHV terminal repeats [21]. We repeated our ChIP-qPCR analysis and additionally performed ChIP-Seq from the bulk-selected cultures at 33 days p.i., a similar late time point as in our previous MHV-68Δ50 or MHV-68Δ50-kLANA experiments.…”

Section: Resultsmentioning

confidence: 72%

“…Given this, we wondered whether unique properties of kLANA that are not conserved in murine homolog (mLANA) may be required for rapid acquisition of H3K27-me3, potentially explaining the absence of this mark from MHV-68 episomes in singly or co-infected cells. Although KSHV and MHV-68 LANA preferentially bind to their cognate sites within the terminal repeats, a number of recent studies have suggested that they can also reciprocally support genome replication and maintenance [21-24]. We therefore sought to investigate the ability of a recombinant MHV-68 genome harboring a kLANA gene to attract H3K27-me3 marks.…”

Section: Resultsmentioning

confidence: 99%

“…We therefore sought to investigate the ability of a recombinant MHV-68 genome harboring a kLANA gene to attract H3K27-me3 marks. For this purpose, we introduced the ORF50 deletion in the background of a chimeric MHV-68 mutant (recently generated by Habison and colleagues [21]) in which mLANA was replaced by kLANA, permitting us to investigate the chromatin landscape of latently replicating MHV-68 genomes that are maintained by kLANA instead of mLANA. As expected, MLE12 cells infected with either the mLANA-expressing MHV-68Δ50 virus or the MHV-68Δ50-kLANA mutant showed readily detectable expression of the respective LANA protein (Fig 6D).…”

Section: Resultsmentioning

confidence: 99%

“…While kLANA has been shown to support episome maintenance of MHV-68 genomes [21-23], it seemed formally possible that it may be selectively defective for PRC recruitment in a non-syngeneic context, for example due to differential binding stoichiometry at the terminal repeats or the lack of low affinity binding sites in the coding region. We therefore also tested whether trans-complementation of a LANA-deleted KSHV genome with either kLANA or mLANA would result in differential behavior towards PRC repression, expecting that mLANA-maintained KSHV genomes may be unable to acquire H3K27-me3 marks.…”

Section: Resultsmentioning

confidence: 99%

See 3 more Smart Citations

A comparative epigenome analysis of gammaherpesviruses suggests cis-acting sequence features as critical mediators of rapid polycomb recruitment

Günther

Fröhlich

Herrde

et al. 2019

Preprint

View full text Add to dashboard Cite

Latent Kaposi sarcoma-associated herpesvirus (KSHV) genomes rapidly acquire distinct patterns of the activating histone modification H3K4-me3 as well as repressive H3K27-me3 marks, a modification linked to transcriptional silencing by polycomb repressive complexes (PRC). Interestingly, PRCs have recently been reported to restrict viral gene expression in a number of other viral systems, suggesting they may play a broader role in controlling viral chromatin. If so, it is an intriguing possibility that latency establishment may result from viral subversion of polycomb-mediated host responses to exogenous DNA.To investigate such scenarios we sought to establish whether rapid repression by PRC constitutes a general hallmark of herpesvirus latency. For this purpose, we performed a comparative epigenome analysis of KSHV and the related murine gammaherpesvirus 68 (MHV-68). We demonstrate that, while latently replicating MHV-68 genomes readily acquire distinct patterns of activation-associated histone modifications upon de novo infection, they fundamentally differ in their ability to efficiently attract H3K27-me3 marks. Statistical analyses of ChIP-seq data from in vitro infected cells as well as in vivo latency reservoirs furthermore suggest that, whereas KSHV rapidly attracts PRCs in a genome-wide manner, H3K27-me3 acquisition by MHV-68 genomes may require spreading from initial seed sites to which PRC are recruited as the result of an inefficient or stochastic recruitment, and that immune pressure may be needed to select for latency pools harboring PRC-silenced episomes in vivo.Using co-infection experiments and recombinant viruses, we also show that KSHV’S ability to rapidly and efficiently acquire H3K27-me3 marks does not depend on the host cell environment or unique properties of the KSHV-encoded LANA protein, but rather requires specific cis-acting sequence features. We show that the non-canonical PRC1.1 component KDM2B, a factor which binds to unmethylated CpG motifs, is efficiently recruited to KSHV genomes, indicating that CpG island characteristics may constitute these features. In accord with the fact that, compared to MHV-68, KSHV genomes exhibit a fundamentally higher density of CpG motifs, we furthermore demonstrate efficient acquisition of H2AK119-ub by KSHV and H3K36-me2 by MHV-68 (but not vice versa), furthermore supporting the notion that KSHV genomes rapidly attract PRC1.1 complexes in a genome-wide fashion. Collectively, our results suggest that rapid PRC silencing is not a universal feature of viral latency, but that some viruses may rather have adopted distinct genomic features to specifically exploit default host pathways that repress epigenetically naive, CpG-rich DNA.Author SummaryDuring herpesvirus latency, viral genomes persists as partially repressed nuclear episomes which do not express genes required for progeny production. Latently infected cells not only form a reservoir of lifelong persistence but also represent the driving force in cancers associated with tumorigenic herpesviruses such as KSHV. Hence, it is fundamentally important to understand the mechanisms controlling latency. We have shown previously that latent KSHV episomes rapidly acquire H3K27-me3, a histone mark associated with polycomb repressive complexes (PRC). PRCs play a pivotal role in the control of developmental genes but are also involved in the pathogenesis of several tumors. We here investigated whether PRC-repression represents a general feature of herpesvirus latency. By performing side-by-side analyses of KSHV and the related MHV-68 we show that the latter indeed has a fundamentally lower propensity to acquire H3K27-me3, and that KSHV’S ability to rapidly attract this mark is most likely the result of a specific sequence composition that promotes recruitment of non-canonical PRC1 (a complex which is important for the regulation of cellular CpG islands). Our results have widespread implications for nuclear DNA viruses and suggest that some viruses have specifically evolved to exploit common host responses to epigenetically naive DNA.

show abstract

Section: Resultsmentioning

confidence: 72%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 2 more Smart Citations

A comparative epigenome analysis of gammaherpesviruses suggests cis-acting sequence features as critical mediators of rapid polycomb recruitment

Günther

Fröhlich

Herrde

et al. 2019

Preprint

View full text Add to dashboard Cite

show abstract

“…A chimera virus was developed (a mouse virus with a human viral gene) that inhibits human LANA protein (essential for maintaining infection and causing cancer) for treatment of human herpes virus infection and its associated cancers. Such strategies can also be applied for the generation of chimera viruses, which can be effective against some other lethal viruses such as the Epstein-Barr virus or the human papilloma virus responsible for cervical cancers (Habison et al, 2017). Very recently, a chimeric antigen receptor T (CAR-T) cell has been developed for the treatment of relapsed or refractory acute lymphoblastic leukemia.…”

Section: Chimeric Enterovirusmentioning

confidence: 99%

Next generation agents (synthetic agents): Emerging threats and challenges in detection, protection, and decontamination

Sharma

Gupta

Ahmad

et al. 2020

Handbook on Biological Warfare Preparedness

View full text Add to dashboard Cite

MicroRNA-181a regulates IFN-γ expression in effector CD8+ T cell differentiation

et al. 2020

Self Cite

View full text Add to dashboard Cite

CD8 + T cells are key players in immunity against intracellular infections and tumors. The main cytokine associated with these protective responses is interferon-γ (IFN-γ), whose production is known to be regulated at the transcriptional level during CD8 + T cell differentiation. Here we found that microRNAs constitute a posttranscriptional brake to IFN-γ expression by CD8 + T cells, since the genetic interference with the Dicer processing machinery resulted in the overproduction of IFN-γ by both thymic and peripheral CD8 + T cells. Using a gene reporter mouse for IFN-γ locus activity, we compared the microRNA repertoires associated with the presence or absence of IFN-γ expression. This allowed us to identify a set of candidates, including miR-181a and miR-451, which were functionally tested in overexpression experiments using synthetic mimics in peripheral CD8 + T cell cultures. We found that miR-181a limits IFN-γ production by suppressing the expression of the transcription factor Id2, which in turn promotes the Ifng expression program. Importantly, upon MuHV-4 challenge, miR-181a-deficient mice showed a more vigorous IFN-γ + CD8 + T cell response and were able to control viral infection significantly more efficiently than control mice. These data collectively establish a novel role for miR-181a in regulating IFN-γ-mediated effector CD8 + T cell responses in vitro and in vivo.

show abstract

Cross-species conservation of episome maintenance provides a basis for in vivo investigation of Kaposi's sarcoma herpesvirus LANA

Cited by 20 publications

References 73 publications

A comparative epigenome analysis of gammaherpesviruses suggests cis-acting sequence features as critical mediators of rapid polycomb recruitment

A comparative epigenome analysis of gammaherpesviruses suggests cis-acting sequence features as critical mediators of rapid polycomb recruitment

Next generation agents (synthetic agents): Emerging threats and challenges in detection, protection, and decontamination

MicroRNA-181a regulates IFN-γ expression in effector CD8+ T cell differentiation

Contact Info

Product

Resources

About