Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2022 |
| Outros Autores: | , , , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
| Texto Completo: | http://hdl.handle.net/10362/134341 |
Resumo: | PD/BD/142851/2018 PD/00065/2013 MIT-EXPL/BIO/0008/2017 IF/00959/2015 |
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Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolinG-quadruplex ligandsMolecular receptorsNucleolinPre-miRNA-149RNA G-quadruplexPharmaceutical SciencePD/BD/142851/2018 PD/00065/2013 MIT-EXPL/BIO/0008/2017 IF/00959/2015One of the most significant challenges in capturing and detecting biomarkers is the choice of an appropriate biomolecular receptor. Recently, RNA G-quadruplexes emerged as plausible receptors due to their ability to recognize with high-affinity proteins. Herein, we have unveiled and characterized the capability of the precursor microRNA 149 to form a G-quadruplex structure and determined the role that some ligands may have in its folding and binding capacity to nucleolin. The G-quadruplex formation was induced by K+ ions and stabilized by ligands, as demonstrated by nuclear magnetic resonance and circular dichroism experiments. Surface plasmon resonance measurements showed a binding affinity of precursor microRNA 149 towards ligands in the micromolar range (10−5–10−6 M) and a strong binding affinity to nucleolin RNA-binding domains 1 and 2 (8.38 × 10−10 M). Even in the presence of the ligand PhenDC3, the binding remains almost identical and in the same order of magnitude (4.46 × 10−10 M). The molecular interactions of the RNA G-quadruplex motif found in precursor miRNA 149 (5′-GGGAGGGAGGGACGGG- 3′) and nucleolin RNA-binding domains 1 and 2 were explored by means of molecular docking and molecular dynamics studies. The results showed that RNA G-quadruplex binds to a cavity between domains 1 and 2 of the protein. Then, complex formation was also evaluated through polyacrylamide gel electrophoresis. The results suggest that precursor microRNA 149/ligands and precursor microRNA 149/nucleolin RNA-binding domains 1 and 2 form stable molecular complexes. The in vitro co-localization of precursor microRNA 149 and nucleolin in PC3 cells was demonstrated using confocal microscopy. Finally, a rapid and straightforward microfluidic strategy was employed to check the ability of precursor microRNA 149 to capture nucleolin RNA-binding domains 1 and 2. The results revealed that precursor microRNA 149 can capture nucleolin RNA-binding domains 1 and 2 labeled with Fluorescein 5-isothiocyanate in a concentration-dependent manner, but PhenDC3 complexation seems to decrease the ability of precursor microRNA 149 to capture the protein. Overall, our results proved the formation of the G-quadruplex structure in the precursor microRNA 149 and the ability to recognize and detect nucleolin. This proof-of-concept study could open up a new framework for developing new strategies to design improved molecular receptors for capture and detection of nucleolin in complex biological samples.UCIBIO - Applied Molecular Biosciences UnitDQ - Departamento de QuímicaRUNSantos, TiagoMiranda, AndréImbert, LionelJardim, AndreiaCaneira, Catarina R. F.Chu, VirgíniaConde, João PedroCampello, Maria Paula CabralPaulo, AntónioSalgado, GilmarCabrita, Eurico J.Cruz, Carla2022-03-11T23:22:16Z2022-02-012022-02-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/134341eng0928-0987PURE: 36467709https://doi.org/10.1016/j.ejps.2021.106093info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T05:12:48Zoai:run.unl.pt:10362/134341Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:48:04.081938Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
| dc.title.none.fl_str_mv |
Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin |
| title |
Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin |
| spellingShingle |
Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin Santos, Tiago G-quadruplex ligands Molecular receptors Nucleolin Pre-miRNA-149 RNA G-quadruplex Pharmaceutical Science |
| title_short |
Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin |
| title_full |
Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin |
| title_fullStr |
Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin |
| title_full_unstemmed |
Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin |
| title_sort |
Pre-miRNA-149 G-quadruplex as a molecular agent to capture nucleolin |
| author |
Santos, Tiago |
| author_facet |
Santos, Tiago Miranda, André Imbert, Lionel Jardim, Andreia Caneira, Catarina R. F. Chu, Virgínia Conde, João Pedro Campello, Maria Paula Cabral Paulo, António Salgado, Gilmar Cabrita, Eurico J. Cruz, Carla |
| author_role |
author |
| author2 |
Miranda, André Imbert, Lionel Jardim, Andreia Caneira, Catarina R. F. Chu, Virgínia Conde, João Pedro Campello, Maria Paula Cabral Paulo, António Salgado, Gilmar Cabrita, Eurico J. Cruz, Carla |
| author2_role |
author author author author author author author author author author author |
| dc.contributor.none.fl_str_mv |
UCIBIO - Applied Molecular Biosciences Unit DQ - Departamento de Química RUN |
| dc.contributor.author.fl_str_mv |
Santos, Tiago Miranda, André Imbert, Lionel Jardim, Andreia Caneira, Catarina R. F. Chu, Virgínia Conde, João Pedro Campello, Maria Paula Cabral Paulo, António Salgado, Gilmar Cabrita, Eurico J. Cruz, Carla |
| dc.subject.por.fl_str_mv |
G-quadruplex ligands Molecular receptors Nucleolin Pre-miRNA-149 RNA G-quadruplex Pharmaceutical Science |
| topic |
G-quadruplex ligands Molecular receptors Nucleolin Pre-miRNA-149 RNA G-quadruplex Pharmaceutical Science |
| description |
PD/BD/142851/2018 PD/00065/2013 MIT-EXPL/BIO/0008/2017 IF/00959/2015 |
| publishDate |
2022 |
| dc.date.none.fl_str_mv |
2022-03-11T23:22:16Z 2022-02-01 2022-02-01T00:00:00Z |
| dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10362/134341 |
| url |
http://hdl.handle.net/10362/134341 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
0928-0987 PURE: 36467709 https://doi.org/10.1016/j.ejps.2021.106093 |
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info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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