Perturbation of m6A Writers Reveals Two Distinct Classes of mRNA Methylation at Internal and 5′ Sites
N6-methyladenosine (m6A) is a common modification of mRNA with potential roles in fine-tuning the RNA life cycle. Here, we identify a dense network of proteins interacting with METTL3, a component of the methyltransferase complex, and show that three of them (WTAP, METTL14, and KIAA1429) are require...
| Main Authors: | , , , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Elsevier
2015
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| Online Access: | http://hdl.handle.net/1721.1/96674 https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0002-7203-4299 https://orcid.org/0000-0002-4227-5163 https://orcid.org/0000-0001-8567-2049 |
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| author | Schwartz, Schraga Mumbach, Maxwell R. Jovanovic, Marko Wang, Tim Maciag, Karolina Bushkin, G. Guy Mertins, Philipp Ter-Ovanesyan, Dmitry Habib, Naomi Cacchiarelli, Davide Sanjana, Neville E. Freinkman, Elizaveta Pacold, Michael E. Satija, Rahul Hacohen, Nir Zhang, Feng Regev, Aviv Mikkelsen, Tarjei Sigurd, 1978- Carr, Steven A Lander, Eric Steven |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Schwartz, Schraga Mumbach, Maxwell R. Jovanovic, Marko Wang, Tim Maciag, Karolina Bushkin, G. Guy Mertins, Philipp Ter-Ovanesyan, Dmitry Habib, Naomi Cacchiarelli, Davide Sanjana, Neville E. Freinkman, Elizaveta Pacold, Michael E. Satija, Rahul Hacohen, Nir Zhang, Feng Regev, Aviv Mikkelsen, Tarjei Sigurd, 1978- Carr, Steven A Lander, Eric Steven |
| author_sort | Schwartz, Schraga |
| collection | MIT |
| description | N6-methyladenosine (m6A) is a common modification of mRNA with potential roles in fine-tuning the RNA life cycle. Here, we identify a dense network of proteins interacting with METTL3, a component of the methyltransferase complex, and show that three of them (WTAP, METTL14, and KIAA1429) are required for methylation. Monitoring m6A levels upon WTAP depletion allowed the definition of accurate and near single-nucleotide resolution methylation maps and their classification into WTAP-dependent and -independent sites. WTAP-dependent sites are located at internal positions in transcripts, topologically static across a variety of systems we surveyed, and inversely correlated with mRNA stability, consistent with a role in establishing “basal” degradation rates. WTAP-independent sites form at the first transcribed base as part of the cap structure and are present at thousands of sites, forming a previously unappreciated layer of transcriptome complexity. Our data shed light on the proteomic and transcriptional underpinnings of this RNA modification. |
| first_indexed | 2024-09-23T09:36:17Z |
| format | Article |
| id | mit-1721.1/96674 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T09:36:17Z |
| publishDate | 2015 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/966742022-09-30T15:36:23Z Perturbation of m6A Writers Reveals Two Distinct Classes of mRNA Methylation at Internal and 5′ Sites Schwartz, Schraga Mumbach, Maxwell R. Jovanovic, Marko Wang, Tim Maciag, Karolina Bushkin, G. Guy Mertins, Philipp Ter-Ovanesyan, Dmitry Habib, Naomi Cacchiarelli, Davide Sanjana, Neville E. Freinkman, Elizaveta Pacold, Michael E. Satija, Rahul Hacohen, Nir Zhang, Feng Regev, Aviv Mikkelsen, Tarjei Sigurd, 1978- Carr, Steven A Lander, Eric Steven Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences Whitehead Institute for Biomedical Research Koch Institute for Integrative Cancer Research at MIT Wang, Tim Zhang, Feng Carr, Steven A. Lander, Eric S. Regev, Aviv N6-methyladenosine (m6A) is a common modification of mRNA with potential roles in fine-tuning the RNA life cycle. Here, we identify a dense network of proteins interacting with METTL3, a component of the methyltransferase complex, and show that three of them (WTAP, METTL14, and KIAA1429) are required for methylation. Monitoring m6A levels upon WTAP depletion allowed the definition of accurate and near single-nucleotide resolution methylation maps and their classification into WTAP-dependent and -independent sites. WTAP-dependent sites are located at internal positions in transcripts, topologically static across a variety of systems we surveyed, and inversely correlated with mRNA stability, consistent with a role in establishing “basal” degradation rates. WTAP-independent sites form at the first transcribed base as part of the cap structure and are present at thousands of sites, forming a previously unappreciated layer of transcriptome complexity. Our data shed light on the proteomic and transcriptional underpinnings of this RNA modification. National Human Genome Research Institute (U.S.). Centers of Excellence in Genomic Science (P50 HG006193) National Human Genome Research Institute (U.S.) (U54 HG003067) National Institutes of Health (U.S.) (Pioneer Award) 2015-04-17T17:28:08Z 2015-04-17T17:28:08Z 2014-06 2014-04 Article http://purl.org/eprint/type/JournalArticle 22111247 http://hdl.handle.net/1721.1/96674 Schwartz, Schraga, Maxwell R. Mumbach, Marko Jovanovic, Tim Wang, Karolina Maciag, G. Guy Bushkin, Philipp Mertins, et al. “Perturbation of m6A Writers Reveals Two Distinct Classes of mRNA Methylation at Internal and 5′ Sites.” Cell Reports 8, no. 1 (July 2014): 284–296. https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0002-7203-4299 https://orcid.org/0000-0002-4227-5163 https://orcid.org/0000-0001-8567-2049 en_US http://dx.doi.org/10.1016/j.celrep.2014.05.048 Cell Reports Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License http://creativecommons.org/licenses/by-nc-nd/3.0/ application/pdf Elsevier Elsevier Open Access |
| spellingShingle | Schwartz, Schraga Mumbach, Maxwell R. Jovanovic, Marko Wang, Tim Maciag, Karolina Bushkin, G. Guy Mertins, Philipp Ter-Ovanesyan, Dmitry Habib, Naomi Cacchiarelli, Davide Sanjana, Neville E. Freinkman, Elizaveta Pacold, Michael E. Satija, Rahul Hacohen, Nir Zhang, Feng Regev, Aviv Mikkelsen, Tarjei Sigurd, 1978- Carr, Steven A Lander, Eric Steven Perturbation of m6A Writers Reveals Two Distinct Classes of mRNA Methylation at Internal and 5′ Sites |
| title | Perturbation of m6A Writers Reveals Two Distinct Classes of mRNA Methylation at Internal and 5′ Sites |
| title_full | Perturbation of m6A Writers Reveals Two Distinct Classes of mRNA Methylation at Internal and 5′ Sites |
| title_fullStr | Perturbation of m6A Writers Reveals Two Distinct Classes of mRNA Methylation at Internal and 5′ Sites |
| title_full_unstemmed | Perturbation of m6A Writers Reveals Two Distinct Classes of mRNA Methylation at Internal and 5′ Sites |
| title_short | Perturbation of m6A Writers Reveals Two Distinct Classes of mRNA Methylation at Internal and 5′ Sites |
| title_sort | perturbation of m6a writers reveals two distinct classes of mrna methylation at internal and 5 sites |
| url | http://hdl.handle.net/1721.1/96674 https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0002-7203-4299 https://orcid.org/0000-0002-4227-5163 https://orcid.org/0000-0001-8567-2049 |
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