RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling
Invading pathogen nucleic acids are recognized and bound by cytoplasmic (retinoic acid-inducible gene I [RIG-I]-like) and membrane-bound (Toll-like) pattern recognition receptors to activate innate immune signaling. Modified nucleotides, when present in RNA molecules, diminish the magnitude of these...
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| Format: | Article |
| Language: | en_US |
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American Society for Microbiology
2017
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| Online Access: | http://hdl.handle.net/1721.1/107388 https://orcid.org/0000-0002-9387-8212 |
| _version_ | 1811081534722015232 |
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| author | Durbin, Ann Fiegen Wang, Chen Marcotrigiano, Joseph Gehrke, Lee |
| author2 | Massachusetts Institute of Technology. Institute for Medical Engineering & Science |
| author_facet | Massachusetts Institute of Technology. Institute for Medical Engineering & Science Durbin, Ann Fiegen Wang, Chen Marcotrigiano, Joseph Gehrke, Lee |
| author_sort | Durbin, Ann Fiegen |
| collection | MIT |
| description | Invading pathogen nucleic acids are recognized and bound by cytoplasmic (retinoic acid-inducible gene I [RIG-I]-like) and membrane-bound (Toll-like) pattern recognition receptors to activate innate immune signaling. Modified nucleotides, when present in RNA molecules, diminish the magnitude of these signaling responses. However, mechanisms explaining the blunted signaling have not been elucidated. In this study, we used several independent biological assays, including inhibition of virus replication, RIG-I:RNA binding assays, and limited trypsin digestion of RIG-I:RNA complexes, to begin to understand how RNAs containing modified nucleotides avoid or suppress innate immune signaling. The experiments were based on a model innate immune activating RNA molecule, the polyU/UC RNA domain of hepatitis C virus, which was transcribed in vitro with canonical nucleotides or with one of eight modified nucleotides. The approach revealed signature assay responses associated with individual modified nucleotides or classes of modified nucleotides. For example, while both N-6-methyladenosine (m6A) and pseudouridine nucleotides correlate with diminished signaling, RNA containing m6A modifications bound RIG-I poorly, while RNA containing pseudouridine bound RIG-I with high affinity but failed to trigger the canonical RIG-I conformational changes associated with robust signaling. These data advance understanding of RNA-mediated innate immune signaling, with additional relevance for applying nucleotide modifications to RNA therapeutics. |
| first_indexed | 2024-09-23T11:48:05Z |
| format | Article |
| id | mit-1721.1/107388 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:48:05Z |
| publishDate | 2017 |
| publisher | American Society for Microbiology |
| record_format | dspace |
| spelling | mit-1721.1/1073882022-10-01T06:08:27Z RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling Durbin, Ann Fiegen Wang, Chen Marcotrigiano, Joseph Gehrke, Lee Massachusetts Institute of Technology. Institute for Medical Engineering & Science Gehrke, Lee Invading pathogen nucleic acids are recognized and bound by cytoplasmic (retinoic acid-inducible gene I [RIG-I]-like) and membrane-bound (Toll-like) pattern recognition receptors to activate innate immune signaling. Modified nucleotides, when present in RNA molecules, diminish the magnitude of these signaling responses. However, mechanisms explaining the blunted signaling have not been elucidated. In this study, we used several independent biological assays, including inhibition of virus replication, RIG-I:RNA binding assays, and limited trypsin digestion of RIG-I:RNA complexes, to begin to understand how RNAs containing modified nucleotides avoid or suppress innate immune signaling. The experiments were based on a model innate immune activating RNA molecule, the polyU/UC RNA domain of hepatitis C virus, which was transcribed in vitro with canonical nucleotides or with one of eight modified nucleotides. The approach revealed signature assay responses associated with individual modified nucleotides or classes of modified nucleotides. For example, while both N-6-methyladenosine (m6A) and pseudouridine nucleotides correlate with diminished signaling, RNA containing m6A modifications bound RIG-I poorly, while RNA containing pseudouridine bound RIG-I with high affinity but failed to trigger the canonical RIG-I conformational changes associated with robust signaling. These data advance understanding of RNA-mediated innate immune signaling, with additional relevance for applying nucleotide modifications to RNA therapeutics. United States. Public Health Service (Grants CA159132 and AI100190) 2017-03-10T19:08:38Z 2017-03-10T19:08:38Z 2016-09 2016-05 Article http://purl.org/eprint/type/JournalArticle 2150-7511 http://hdl.handle.net/1721.1/107388 Durbin, Ann Fiegen et al. “RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling.” mBio 7.5 (2016): e00833-16. https://orcid.org/0000-0002-9387-8212 en_US http://dx.doi.org/10.1128/mBio.00833-16 mBio Creative Commons Attribution 4.0 International License http://creativecommons.org/licenses/by/4.0/ application/pdf American Society for Microbiology American Society for Microbiology |
| spellingShingle | Durbin, Ann Fiegen Wang, Chen Marcotrigiano, Joseph Gehrke, Lee RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling |
| title | RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling |
| title_full | RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling |
| title_fullStr | RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling |
| title_full_unstemmed | RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling |
| title_short | RNAs Containing Modified Nucleotides Fail To Trigger RIG-I Conformational Changes for Innate Immune Signaling |
| title_sort | rnas containing modified nucleotides fail to trigger rig i conformational changes for innate immune signaling |
| url | http://hdl.handle.net/1721.1/107388 https://orcid.org/0000-0002-9387-8212 |
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