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High Throughput In Vitro Assessment of Latency Reversing Agents on HIV Transcription and Splicing
Published on: January 22, 2019
Gene silencing in HIV-1 latency by polycomb repressive group
Hyeon Guk Kim1, Kyung-Chang Kim, Tae-Young Roh
1Division of AIDS, Center for Immunology and Pathology, Korea National Institute of Health, Chung-buk, Republic of Korea.
Epigenetic modifications, including histone H3K27 trimethylation and H2A ubiquitylation, are implicated in human immunodeficiency virus-1 (HIV-1) latency. These changes in NCHA cells suggest a role for polycomb group proteins in HIV-1 persistence.
Area of Science:
- Virology
- Epigenetics
- Immunology
Background:
- Human immunodeficiency virus-1 (HIV-1) latency in CD4+ T cells creates persistent reservoirs, hindering eradication.
- The precise molecular mechanisms driving HIV-1 latency remain incompletely understood.
Purpose of the Study:
- To investigate the role of histone modifications in HIV-1 latency.
- To analyze epigenetic changes in novel NCHA cells latently infected with HIV-1.
Main Methods:
- Characterized expression of core histones and histone deacetylases (HDACs) using RT-PCR, ELISA, and western blot.
- Analyzed histone H3 Lys9 (H3K9) and Lys27 (H3K27) acetylation and methylation via western blot and ChIP-seq.
- Assessed H2A ubiquitylation levels.
Main Results:
- Core histone and HDAC expression showed no significant differences between NCHA and parental cells.
- Histone H3K9 and H3K27 acetylation levels were similar; however, di- and tri-methylation increased in NCHA cells.
- Upregulation of PRC2 (EED) and PRC1 (BMI1, RING2) components and increased H2A ubiquitylation were observed in NCHA cells.
Conclusions:
- Histone H3K27 trimethylation and H2A ubiquitylation, mediated by polycomb group proteins, are crucial for epigenetic silencing in HIV-1 latency.
- These epigenetic modifications contribute to the establishment and maintenance of HIV-1 latent reservoirs in NCHA cells.
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