Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II
Argonaute (Ago) proteins mediate posttranscriptional gene repression by binding guide miRNAs to regulate targeted RNAs. To confidently assess Ago-bound small RNAs, we adapted a mouse embryonic stem cell system to express a single epitope-tagged Ago protein family member in an inducible manner. Here,...
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| Format: | Article |
| Language: | en_US |
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Elsevier
2017
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| Online Access: | http://hdl.handle.net/1721.1/110327 https://orcid.org/0000-0002-3621-0144 https://orcid.org/0000-0003-1465-1691 |
| _version_ | 1826207046284869632 |
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| author | Zamudio, Jesse Ray Kelly, Timothy James Sharp, Phillip A. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Zamudio, Jesse Ray Kelly, Timothy James Sharp, Phillip A. |
| author_sort | Zamudio, Jesse Ray |
| collection | MIT |
| description | Argonaute (Ago) proteins mediate posttranscriptional gene repression by binding guide miRNAs to regulate targeted RNAs. To confidently assess Ago-bound small RNAs, we adapted a mouse embryonic stem cell system to express a single epitope-tagged Ago protein family member in an inducible manner. Here, we report the small RNA profile of Ago-deficient cells and show that Ago-dependent stability is a common feature of mammalian miRNAs. Using this criteria and immunopurification, we identified an Ago-dependent class of noncanonical miRNAs derived from protein-coding gene promoters, which we name transcriptional start site miRNAs (TSS-miRNAs). A subset of promoter-proximal RNA polymerase II (RNAPII) complexes produces hairpin RNAs that are processed in a DiGeorge syndrome critical region gene 8 (Dgcr8)/Drosha-independent but Dicer-dependent manner. TSS-miRNA activity is detectable from endogenous levels and following overexpression of mRNA constructs. Finally, we present evidence of differential expression and conservation in humans, suggesting important roles in gene regulation. |
| first_indexed | 2024-09-23T13:42:39Z |
| format | Article |
| id | mit-1721.1/110327 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T13:42:39Z |
| publishDate | 2017 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1103272022-10-01T16:43:17Z Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II Zamudio, Jesse Ray Kelly, Timothy James Sharp, Phillip A. Massachusetts Institute of Technology. Department of Biology Koch Institute for Integrative Cancer Research at MIT Zamudio, Jesse Ray Kelly, Timothy James Sharp, Phillip A. Argonaute (Ago) proteins mediate posttranscriptional gene repression by binding guide miRNAs to regulate targeted RNAs. To confidently assess Ago-bound small RNAs, we adapted a mouse embryonic stem cell system to express a single epitope-tagged Ago protein family member in an inducible manner. Here, we report the small RNA profile of Ago-deficient cells and show that Ago-dependent stability is a common feature of mammalian miRNAs. Using this criteria and immunopurification, we identified an Ago-dependent class of noncanonical miRNAs derived from protein-coding gene promoters, which we name transcriptional start site miRNAs (TSS-miRNAs). A subset of promoter-proximal RNA polymerase II (RNAPII) complexes produces hairpin RNAs that are processed in a DiGeorge syndrome critical region gene 8 (Dgcr8)/Drosha-independent but Dicer-dependent manner. TSS-miRNA activity is detectable from endogenous levels and following overexpression of mRNA constructs. Finally, we present evidence of differential expression and conservation in humans, suggesting important roles in gene regulation. United States. Public Health Service (grant RO1 GM34277) National Cancer Institute (U.S.) (PO1-CA42063) National Cancer Institute (U.S.) (Koch Institute Support (core) grant P30-CA14051) National Institutes of Health (U.S.). Ruth L. Kirschstein National Research Service Award (F32GM101872) National Institutes of Health (U.S.). Ruth L. Kirschstein National Research Service Award (F32CA139902) 2017-06-27T18:42:04Z 2017-06-27T18:42:04Z 2014-02 2013-11 Article http://purl.org/eprint/type/JournalArticle 0092-8674 1097-4172 http://hdl.handle.net/1721.1/110327 Zamudio, Jesse R., Timothy J. Kelly, and Phillip A. Sharp. “Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II.” Cell 156.5 (2014): 920–934. https://orcid.org/0000-0002-3621-0144 https://orcid.org/0000-0003-1465-1691 en_US http://dx.doi.org/10.1016/j.cell.2014.01.041 Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Zamudio, Jesse Ray Kelly, Timothy James Sharp, Phillip A. Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II |
| title | Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II |
| title_full | Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II |
| title_fullStr | Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II |
| title_full_unstemmed | Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II |
| title_short | Argonaute-Bound Small RNAs from Promoter-Proximal RNA Polymerase II |
| title_sort | argonaute bound small rnas from promoter proximal rna polymerase ii |
| url | http://hdl.handle.net/1721.1/110327 https://orcid.org/0000-0002-3621-0144 https://orcid.org/0000-0003-1465-1691 |
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