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Beyond the binding site: In vivo Identification of tbx2, smarca5 and wnt5b as molecular targets of CNBP during embryonic development

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dc.citation.titlePLoS ONE
dc.citation.volume8
dc.creatorArmas, Pablo
dc.creatorMargarit, Ezequiel
dc.creatorMouguelar, Valeria
dc.creatorAllende, Miguel L.
dc.creatorCalcaterra, Nora B.
dc.date.accessioned2024-06-25T15:14:36Z
dc.date.available2024-06-25T15:14:36Z
dc.date.issued2013-05-07
dc.description.abstractCNBP is a nucleic acid chaperone implicated in vertebrate craniofacial development, as well as in myotonic dystrophy type 2 (DM2) and sporadic inclusion body myositis (sIBM) human muscle diseases. CNBP is highly conserved among vertebrates and has been implicated in transcriptional regulation; however, its DNA binding sites and molecular targets remain elusive. The main goal of this work was to identify CNBP DNA binding sites that might reveal target genes involved in vertebrate embryonic development. To accomplish this, we used a recently described yeast one-hybrid assay to identify DNA sequences bound in vivo by CNBP. Bioinformatic analyses revealed that these sequences are G-enriched and show high frequency of putative G-quadruplex DNA secondary structure. Moreover, an in silico approach enabled us to establish the CNBP DNA-binding site and to predict CNBP putative targets based on gene ontology terms and synexpression with CNBP. The direct interaction between CNBP and candidate genes was proved by EMSA and ChIP assays. Besides, the role of CNBP upon the identified genes was validated in loss-of-function experiments in developing zebrafish. We successfully confirmed that CNBP up-regulates tbx2b and smarca5, and down-regulates wnt5b gene expression. The highly stringent strategy used in this work allowed us to identify new CNBP target genes functionally important in different contexts of vertebrate embryonic development. Furthermore, it represents a novel approach toward understanding the biological function and regulatory networks involving CNBP in the biology of vertebrates.
dc.description.filFil: Armas, Pablo. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología Molecular y Celular de Rosario (CONICET-IBR); Argentina.
dc.description.filFil: Margarit, Ezequiel. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología Molecular y Celular de Rosario (CONICET-IBR); Argentina.
dc.description.filFil: Mouguelar, Valeria. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología Molecular y Celular de Rosario (CONICET-IBR); Argentina.
dc.description.filFil: Allende, Miguel L. Universidad de Chile. Facultad de Ciencias. FONDAP Center for Genome Regulation; Santiago, Chile.
dc.description.filFil: Calcaterra, Nora B. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología Molecular y Celular de Rosario (CONICET-IBR); Argentina.
dc.description.sponsorshipFondo de Financiamiento de Centros de Investigación en Áreas Prioritarias (FONDAP): 15090007
dc.description.sponsorshipInternational Centre For Genetic Engineering And Biotechnology (ICGEB): CRP/CHI11-01
dc.description.sponsorshipAgencia Nacional de Promoción Científica y Tecnológica (ANPCyT): PICT 07-00648, PICT 11-1540
dc.description.sponsorshipConsejo Nacional de Investigaciones Científicas y Técnicas (CONICET): PIP 00480 and PIP 00773
dc.description.versionpeerreviewed
dc.format.extent1-12
dc.identifier.issn1932-6203
dc.identifier.urihttps://hdl.handle.net/2133/27343
dc.language.isoen
dc.publisherPublic Library of Science
dc.relation.publisherversionhttps://journals.plos.org/plosone/article?id=10.1371/journal.pone.0063234
dc.relation.publisherversionhttps://doi.org/10.1371/journal.pone.0063234
dc.rightsopenAccess
dc.rights.holderArmas, Pablo
dc.rights.holderMargarit, Ezequiel
dc.rights.holderMouguelar, Valeria
dc.rights.holderAllende, Miguel L.
dc.rights.holderCalcaterra, Nora B.
dc.rights.holderUniversidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas
dc.rights.textAttribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectCnbp
dc.subjectMolecular targets
dc.subjectEmbryonic development
dc.titleBeyond the binding site: In vivo Identification of tbx2, smarca5 and wnt5b as molecular targets of CNBP during embryonic development
dc.typearticulo
dc.type.collectionarticulo
dc.type.versionpublishedVersion

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