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| Status |
Public on Mar 17, 2022 |
| Title |
Integrated analyses reveal CHD4 mediating establishment and maintenance of latent KSHV chromatin [CUT&RUN] |
| Organisms |
Homo sapiens; Human gammaherpesvirus 8 |
| Experiment type |
Genome binding/occupancy profiling by high throughput sequencing
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| Summary |
lncRNAs regulate protein functions via formation of protein-RNA complexes. Previous studies have shown that expression of viral lncRNA, polyadenylated nuclear RNA (PAN RNA) is essential for inducible viral genomic looping and distal gene activation during Kaposi's sarcoma-associated herpesvirus (KSHV) reactivation. Here we show an underlying molecular mechanism and regulation of KSHV latency by a viral lncRNA-CHD4 (chromodomain helicase DNA binding protein 4) interaction. Knock-out of the viral RNA binding protein, ORF57 protein, leads to decreased inducible and static viral genomic loops in latent chromatin and a failure to form RNA polymerase II aggregates in the nucleus during reactivation. We identified that CHD4's enzymatic activity silences viral gene expression by preventing nuclear aggregate formation. Furthermore, integrated genomic and proteomic studies together show that KSHV episomes frequently tether near the host cell centromeres and colocalize with a CHD4 protein complex, ChAHP. KSHV episomes detached from these sites when reactivation is triggered, and PAN RNA binds and inhibits CHD4 DNA binding in vitro. Our studies suggest that CHD4 exhibits strong repressor function by preventing inducible enhancer-promoter looping, and is therefore important for the ability of KSHV to establish and maintain latency in a “poised” state at specific host genomic loci.
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| Overall design |
Examination of genome-wide binding of host and viral proteins on the human and KSHV genome using CUT&RUN. Experiments were performed in KSHV naturally infected human primary effusion lymphoma (PEL) cell line BCBL-1 and wild type or ORF57-Stop recombinant KSHV infected iSLK cells.
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| Contributor(s) |
Izumiya Y, Campell M, Kumar A, Lyu Y, Yanagihashi Y, Chantarasrivong C, Nakano K, Davis RR, Tepper CG |
| Citation(s) |
35545047 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| P30 CA093373 |
Cancer Center Support Grant P30 |
UNIVERSITY OF CALIFORNIA DAVIS |
PRIMO N. LARA |
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| Submission date |
Dec 13, 2020 |
| Last update date |
Jun 16, 2022 |
| Contact name |
Clifford G. Tepper |
| E-mail(s) |
cgtepper@ucdavis.edu
|
| Phone |
916-734-7195
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| Organization name |
UC Davis School of Medicine
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| Department |
Biochemistry and Molecular Medicine
|
| Street address |
4645 2nd Avenue, Room 2300A
|
| City |
Sacramento |
| State/province |
CA |
| ZIP/Postal code |
95817 |
| Country |
USA |
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| Platforms (1) |
| GPL25719 |
Illumina HiSeq 4000 (Homo sapiens; Human gammaherpesvirus 8) |
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| Samples (30)
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| This SubSeries is part of SuperSeries: |
| GSE163695 |
Integrated analyses reveal CHD4 mediating establishment and maintenance of latent KSHV chromatin |
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| Relations |
| BioProject |
PRJNA684898 |
| SRA |
SRP297785 |
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