Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9
TLR9 recognizes unmethylated CpG-containing DNA commonly found in bacteria. Synthetic oligonucleotides containing CpG-motifs (CpG ODNs) recapitulate the activation of TLR9 by microbial DNA, whereas inversion of the CG dinucleotide within the CpG motif to GC (GpC ODNs) renders such ODNs inactive. Thi...
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| Format: | Article |
| Language: | en_US |
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Wiley Blackwell
2014
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| Online Access: | http://hdl.handle.net/1721.1/84991 https://orcid.org/0000-0002-1090-6071 |
| _version_ | 1811093450267820032 |
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| author | Avalos, Ana M. Ploegh, Hidde |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Avalos, Ana M. Ploegh, Hidde |
| author_sort | Avalos, Ana M. |
| collection | MIT |
| description | TLR9 recognizes unmethylated CpG-containing DNA commonly found in bacteria. Synthetic oligonucleotides containing CpG-motifs (CpG ODNs) recapitulate the activation of TLR9 by microbial DNA, whereas inversion of the CG dinucleotide within the CpG motif to GC (GpC ODNs) renders such ODNs inactive. This difference cannot be attributed to binding of ODNs to the full-length TLR9 ectodomain, as both CpG and GpC ODNs bind comparably. Activation of murine TLR9 requires cleavage into an active C-terminal fragment, which binds CpG robustly. We therefore compared the ability of CpG and GpC ODNs to bind to full-length and C-terminal TLR9, and their impact on the cleavage of TLR9. We found that CpG binds better to C-terminal TLR9 when compared with GpC, despite comparably low binding of both ODNs to full-length TLR9. Neither CpG nor GpC ODNs affected TLR9 cleavage in murine RAW 264.7 cells stably expressing TLR9-Myc. Inhibitory ODNs (IN-ODNs) block TLR9 signaling, but how they do so remains unclear. We show here that inhibitory ODNs do not impede TLR9 cleavage but bind to C-terminal TLR9 preferentially, and thereby compete for CpG ODN binding both in RAW cells and in TLR9-deficient cells transduced with TLR9-Myc. Ligand binding to C-terminal fragment thus determines the outcome of activation through TLR9. |
| first_indexed | 2024-09-23T15:45:21Z |
| format | Article |
| id | mit-1721.1/84991 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:45:21Z |
| publishDate | 2014 |
| publisher | Wiley Blackwell |
| record_format | dspace |
| spelling | mit-1721.1/849912022-10-02T03:52:26Z Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9 Avalos, Ana M. Ploegh, Hidde Massachusetts Institute of Technology. Department of Biology Whitehead Institute for Biomedical Research Ploegh, Hidde TLR9 recognizes unmethylated CpG-containing DNA commonly found in bacteria. Synthetic oligonucleotides containing CpG-motifs (CpG ODNs) recapitulate the activation of TLR9 by microbial DNA, whereas inversion of the CG dinucleotide within the CpG motif to GC (GpC ODNs) renders such ODNs inactive. This difference cannot be attributed to binding of ODNs to the full-length TLR9 ectodomain, as both CpG and GpC ODNs bind comparably. Activation of murine TLR9 requires cleavage into an active C-terminal fragment, which binds CpG robustly. We therefore compared the ability of CpG and GpC ODNs to bind to full-length and C-terminal TLR9, and their impact on the cleavage of TLR9. We found that CpG binds better to C-terminal TLR9 when compared with GpC, despite comparably low binding of both ODNs to full-length TLR9. Neither CpG nor GpC ODNs affected TLR9 cleavage in murine RAW 264.7 cells stably expressing TLR9-Myc. Inhibitory ODNs (IN-ODNs) block TLR9 signaling, but how they do so remains unclear. We show here that inhibitory ODNs do not impede TLR9 cleavage but bind to C-terminal TLR9 preferentially, and thereby compete for CpG ODN binding both in RAW cells and in TLR9-deficient cells transduced with TLR9-Myc. Ligand binding to C-terminal fragment thus determines the outcome of activation through TLR9. National Institutes of Health (U.S.) (Grant 5R01AI033456-21) 2014-02-18T20:23:33Z 2014-02-18T20:23:33Z 2011-10 2011-07 Article http://purl.org/eprint/type/JournalArticle 00142980 1521-4141 http://hdl.handle.net/1721.1/84991 Avalos, Ana M., and Hidde L. Ploegh. “Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9.” European Journal of Immunology 41, no. 10 (October 6, 2011): 2820-2827. https://orcid.org/0000-0002-1090-6071 en_US http://dx.doi.org/10.1002/eji.201141563 European Journal of Immunology Creative Commons Attribution-Noncommercial-Share Alike http://creativecommons.org/licenses/by-nc-sa/4.0/ application/pdf Wiley Blackwell PMC |
| spellingShingle | Avalos, Ana M. Ploegh, Hidde Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9 |
| title | Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9 |
| title_full | Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9 |
| title_fullStr | Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9 |
| title_full_unstemmed | Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9 |
| title_short | Competition by inhibitory oligonucleotides prevents binding of CpG to C-terminal TLR9 |
| title_sort | competition by inhibitory oligonucleotides prevents binding of cpg to c terminal tlr9 |
| url | http://hdl.handle.net/1721.1/84991 https://orcid.org/0000-0002-1090-6071 |
| work_keys_str_mv | AT avalosanam competitionbyinhibitoryoligonucleotidespreventsbindingofcpgtocterminaltlr9 AT ploeghhidde competitionbyinhibitoryoligonucleotidespreventsbindingofcpgtocterminaltlr9 |