A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression
Herpes simplex virus (HSV) utilizes and subverts host chromatin mechanisms to express its lytic gene products in mammalian cells. The host cell attempts to silence the incoming viral genome by epigenetic mechanisms, but the viral VP16 and ICP0 proteins promote active chromatin on the viral genome by...
| Main Authors: | , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Language: | en_US |
| Published: |
American Society for Microbiology
2014
|
| Online Access: | http://hdl.handle.net/1721.1/89646 https://orcid.org/0000-0002-6199-6855 |
| _version_ | 1811075254158622720 |
|---|---|
| author | Oh, Hyung Suk Bryant, Kevin F. Nieland, Thomas J. Mazumder, Aprotim Bagul, Mukta Bathe, Mark Root, David E. Knipe, David M. |
| author2 | Massachusetts Institute of Technology. Center for Environmental Health Sciences |
| author_facet | Massachusetts Institute of Technology. Center for Environmental Health Sciences Oh, Hyung Suk Bryant, Kevin F. Nieland, Thomas J. Mazumder, Aprotim Bagul, Mukta Bathe, Mark Root, David E. Knipe, David M. |
| author_sort | Oh, Hyung Suk |
| collection | MIT |
| description | Herpes simplex virus (HSV) utilizes and subverts host chromatin mechanisms to express its lytic gene products in mammalian cells. The host cell attempts to silence the incoming viral genome by epigenetic mechanisms, but the viral VP16 and ICP0 proteins promote active chromatin on the viral genome by recruiting other host epigenetic factors. However, the dependence on VP16 and ICP0 differs in different cell lines, implying cell type-dependent functional contributions of epigenetic factors for HSV gene expression. In this study, we performed a targeted RNA interference (RNAi) screen for cellular chromatin factors that are involved in regulation of herpes simplex virus (HSV) gene expression in U2OS osteosarcoma cells, a cell line that complements ICP0 mutant and VP16 mutant virus replication. In this screen, we found the same general classes of chromatin factors that regulate HSV gene expression in U2OS cells as in other cell types, including histone demethylases (HDMs), histone deacetylases (HDACs), histone acetyltransferases (HATs), and chromatin-remodeling factors, but the specific factors within these classes are different from those identified previously for other cell types. For example, KDM3A and KDM1A (LSD1) both demethylate mono- and dimethylated H3K9, but KDM3A emerged in our screen of U2OS cells. Further, small interfering RNA (siRNA) and inhibitor studies support the idea that KDM1A is more critical in HeLa cells, as observed previously, while KDM3A is more critical in U2OS cells. These results argue that different cellular chromatin factors are critical in different cell lines to carry out the positive and negative epigenetic effects exerted on the HSV genome.
IMPORTANCE Upon entry into the host cell nucleus, the herpes simplex virus genome is subjected to host epigenetic silencing mechanisms. Viral proteins recruit cellular epigenetic activator proteins to reverse and counter the cellular silencing mechanisms. Some of the host silencing and activator functions involved in HSV gene expression have been identified, but there have been indications that the host cell factors may vary in different cell types. In this study, we performed a screen of chromatin factors involved in HSV gene regulation in osteosarcoma cells, and we found that the chromatin factors that are critical for HSV gene expression in these cells are different from those for previously studied cell types. These results argue that the specific chromatin factors operative in different cell lines and cell types may differ. This has implications for epigenetic drugs that are under development. |
| first_indexed | 2024-09-23T10:03:10Z |
| format | Article |
| id | mit-1721.1/89646 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T10:03:10Z |
| publishDate | 2014 |
| publisher | American Society for Microbiology |
| record_format | dspace |
| spelling | mit-1721.1/896462022-09-30T18:34:55Z A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression Oh, Hyung Suk Bryant, Kevin F. Nieland, Thomas J. Mazumder, Aprotim Bagul, Mukta Bathe, Mark Root, David E. Knipe, David M. Massachusetts Institute of Technology. Center for Environmental Health Sciences Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Laboratory for Computational Cell Biology & Biophysics Mazumder, Aprotim Bathe, Mark Herpes simplex virus (HSV) utilizes and subverts host chromatin mechanisms to express its lytic gene products in mammalian cells. The host cell attempts to silence the incoming viral genome by epigenetic mechanisms, but the viral VP16 and ICP0 proteins promote active chromatin on the viral genome by recruiting other host epigenetic factors. However, the dependence on VP16 and ICP0 differs in different cell lines, implying cell type-dependent functional contributions of epigenetic factors for HSV gene expression. In this study, we performed a targeted RNA interference (RNAi) screen for cellular chromatin factors that are involved in regulation of herpes simplex virus (HSV) gene expression in U2OS osteosarcoma cells, a cell line that complements ICP0 mutant and VP16 mutant virus replication. In this screen, we found the same general classes of chromatin factors that regulate HSV gene expression in U2OS cells as in other cell types, including histone demethylases (HDMs), histone deacetylases (HDACs), histone acetyltransferases (HATs), and chromatin-remodeling factors, but the specific factors within these classes are different from those identified previously for other cell types. For example, KDM3A and KDM1A (LSD1) both demethylate mono- and dimethylated H3K9, but KDM3A emerged in our screen of U2OS cells. Further, small interfering RNA (siRNA) and inhibitor studies support the idea that KDM1A is more critical in HeLa cells, as observed previously, while KDM3A is more critical in U2OS cells. These results argue that different cellular chromatin factors are critical in different cell lines to carry out the positive and negative epigenetic effects exerted on the HSV genome. IMPORTANCE Upon entry into the host cell nucleus, the herpes simplex virus genome is subjected to host epigenetic silencing mechanisms. Viral proteins recruit cellular epigenetic activator proteins to reverse and counter the cellular silencing mechanisms. Some of the host silencing and activator functions involved in HSV gene expression have been identified, but there have been indications that the host cell factors may vary in different cell types. In this study, we performed a screen of chromatin factors involved in HSV gene regulation in osteosarcoma cells, and we found that the chromatin factors that are critical for HSV gene expression in these cells are different from those for previously studied cell types. These results argue that the specific chromatin factors operative in different cell lines and cell types may differ. This has implications for epigenetic drugs that are under development. Broad Institute of MIT and Harvard (SPARC grant) Massachusetts Institute of Technology (Faculty Startup funds) Massachusetts Institute of Technology (Samuel A. Goldblith Career Development Professorship) MIT Computational and Systems Biology Initiative (CSBi Merck-MIT postdoctoral fellowship) Merck Research Laboratories (Merck/CSBi Fellowship) Massachusetts Institute of Technology. Center for Environmental Health Sciences (CEHS center grant NIEHS P30-ES002109) National Institutes of Health (U.S.) (grant AI063106) 2014-09-16T15:59:08Z 2014-09-16T15:59:08Z 2014-02 2013-12 Article http://purl.org/eprint/type/JournalArticle 2150-7511 http://hdl.handle.net/1721.1/89646 Oh, H. S., K. F. Bryant, T. J. F. Nieland, A. Mazumder, M. Bagul, M. Bathe, D. E. Root, and D. M. Knipe. “A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression.” mBio 5, no. 1 (December 31, 2013): e01086–13–e01086–13. https://orcid.org/0000-0002-6199-6855 en_US http://dx.doi.org/10.1128/mBio.01086-13 mBio Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/3.0 application/pdf American Society for Microbiology American Society for Microbiology |
| spellingShingle | Oh, Hyung Suk Bryant, Kevin F. Nieland, Thomas J. Mazumder, Aprotim Bagul, Mukta Bathe, Mark Root, David E. Knipe, David M. A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression |
| title | A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression |
| title_full | A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression |
| title_fullStr | A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression |
| title_full_unstemmed | A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression |
| title_short | A Targeted RNA Interference Screen Reveals Novel Epigenetic Factors That Regulate Herpesviral Gene Expression |
| title_sort | targeted rna interference screen reveals novel epigenetic factors that regulate herpesviral gene expression |
| url | http://hdl.handle.net/1721.1/89646 https://orcid.org/0000-0002-6199-6855 |
| work_keys_str_mv | AT ohhyungsuk atargetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT bryantkevinf atargetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT nielandthomasj atargetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT mazumderaprotim atargetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT bagulmukta atargetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT bathemark atargetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT rootdavide atargetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT knipedavidm atargetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT ohhyungsuk targetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT bryantkevinf targetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT nielandthomasj targetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT mazumderaprotim targetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT bagulmukta targetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT bathemark targetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT rootdavide targetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression AT knipedavidm targetedrnainterferencescreenrevealsnovelepigeneticfactorsthatregulateherpesviralgeneexpression |