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Release of human cytomegalovirus from latency by a KAP1/TRIM28 phosphorylation switch.

Rauwel B, Jang SM, Cassano M, Kapopoulou A, Barde I, Trono D - Elife (2015)

Bottom Line: Here, we reveal that HCMV latency in human CD34(+) HSC reflects the recruitment on the viral genome of KAP1, a master co-repressor, together with HP1 and the SETDB1 histone methyltransferase, which results in transcriptional silencing.Correspondingly, HCMV can be forced out of latency by KAP1 knockdown or pharmacological induction of KAP1 phosphorylation, and this process can be potentiated by activating NFkB with TNF-α.These results suggest new approaches both to curtail CMV infection and to purge the virus from organ transplants.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.

ABSTRACT
Human cytomegalovirus (HCMV) is a highly prevalent pathogen that induces life-long infections notably through the establishment of latency in hematopoietic stem cells (HSC). Bouts of reactivation are normally controlled by the immune system, but can be fatal in immuno-compromised individuals such as organ transplant recipients. Here, we reveal that HCMV latency in human CD34(+) HSC reflects the recruitment on the viral genome of KAP1, a master co-repressor, together with HP1 and the SETDB1 histone methyltransferase, which results in transcriptional silencing. During lytic infection, KAP1 is still associated with the viral genome, but its heterochromatin-inducing activity is suppressed by mTOR-mediated phosphorylation. Correspondingly, HCMV can be forced out of latency by KAP1 knockdown or pharmacological induction of KAP1 phosphorylation, and this process can be potentiated by activating NFkB with TNF-α. These results suggest new approaches both to curtail CMV infection and to purge the virus from organ transplants.

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Related in: MedlinePlus

DOI:http://dx.doi.org/10.7554/eLife.06068.037
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fig10: DOI:http://dx.doi.org/10.7554/eLife.06068.037

Mentions: All statistics and Datasets of the ChIP-Seq will be available on Geo as described in Material and methods. We had 163 Million total reads and mapped 135 Million of them on the human genome. Besides this, 400,000 reads corresponded to the HCMV genome. As the CMV-specific signal was broad, the MACS software was not appropriate for peak calling, requesting the development of a special algorithm. All positive, poorly enriched and negative regions were also analyzed by qPCR, which confirmed the results of the ChIP-Seq. The method of ChIP-PCR normalization is explained in the Material and methods section. The Quality control for the ChIP-seq is provided in Author response image 1, together with its Total Input Quality control in Author response image 2:10.7554/eLife.06068.036Author response image 1.


Release of human cytomegalovirus from latency by a KAP1/TRIM28 phosphorylation switch.

Rauwel B, Jang SM, Cassano M, Kapopoulou A, Barde I, Trono D - Elife (2015)

DOI:http://dx.doi.org/10.7554/eLife.06068.037
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4384640&req=5

fig10: DOI:http://dx.doi.org/10.7554/eLife.06068.037
Mentions: All statistics and Datasets of the ChIP-Seq will be available on Geo as described in Material and methods. We had 163 Million total reads and mapped 135 Million of them on the human genome. Besides this, 400,000 reads corresponded to the HCMV genome. As the CMV-specific signal was broad, the MACS software was not appropriate for peak calling, requesting the development of a special algorithm. All positive, poorly enriched and negative regions were also analyzed by qPCR, which confirmed the results of the ChIP-Seq. The method of ChIP-PCR normalization is explained in the Material and methods section. The Quality control for the ChIP-seq is provided in Author response image 1, together with its Total Input Quality control in Author response image 2:10.7554/eLife.06068.036Author response image 1.

Bottom Line: Here, we reveal that HCMV latency in human CD34(+) HSC reflects the recruitment on the viral genome of KAP1, a master co-repressor, together with HP1 and the SETDB1 histone methyltransferase, which results in transcriptional silencing.Correspondingly, HCMV can be forced out of latency by KAP1 knockdown or pharmacological induction of KAP1 phosphorylation, and this process can be potentiated by activating NFkB with TNF-α.These results suggest new approaches both to curtail CMV infection and to purge the virus from organ transplants.

View Article: PubMed Central - PubMed

Affiliation: School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne, Lausanne, Switzerland.

ABSTRACT
Human cytomegalovirus (HCMV) is a highly prevalent pathogen that induces life-long infections notably through the establishment of latency in hematopoietic stem cells (HSC). Bouts of reactivation are normally controlled by the immune system, but can be fatal in immuno-compromised individuals such as organ transplant recipients. Here, we reveal that HCMV latency in human CD34(+) HSC reflects the recruitment on the viral genome of KAP1, a master co-repressor, together with HP1 and the SETDB1 histone methyltransferase, which results in transcriptional silencing. During lytic infection, KAP1 is still associated with the viral genome, but its heterochromatin-inducing activity is suppressed by mTOR-mediated phosphorylation. Correspondingly, HCMV can be forced out of latency by KAP1 knockdown or pharmacological induction of KAP1 phosphorylation, and this process can be potentiated by activating NFkB with TNF-α. These results suggest new approaches both to curtail CMV infection and to purge the virus from organ transplants.

Show MeSH
Related in: MedlinePlus