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http://hdl.handle.net/2445/178642
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DC Field | Value | Language |
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dc.contributor.author | Bueno Martínez, Elena | - |
dc.contributor.author | Sanoguera Miralles, Lara | - |
dc.contributor.author | Valenzuela Palomo, Alberto | - |
dc.contributor.author | Lorca, Víctor | - |
dc.contributor.author | Gómez Sanz, Alicia | - |
dc.contributor.author | Carvalho, Sara | - |
dc.contributor.author | Allen, Jamie | - |
dc.contributor.author | Infante, Mar | - |
dc.contributor.author | Pérez Segura, Pedro | - |
dc.contributor.author | Lázaro García, Conxi | - |
dc.contributor.author | Easton, Douglas F. | - |
dc.contributor.author | Devilee, Peter | - |
dc.contributor.author | Vreeswijk, Maaike P. G. | - |
dc.contributor.author | Hoya, Miguel de la | - |
dc.contributor.author | Velasco, Eladio A. | - |
dc.date.accessioned | 2021-06-25T10:46:24Z | - |
dc.date.available | 2021-06-25T10:46:24Z | - |
dc.date.issued | 2021-06-07 | - |
dc.identifier.uri | http://hdl.handle.net/2445/178642 | - |
dc.description.abstract | RAD51D loss-of-function variants increase lifetime risk of breast and ovarian cancer. Splicing disruption is a frequent pathogenic mechanism associated with variants in susceptibility genes. Herein, we have assessed the splicing and clinical impact of splice-site and exonic splicing enhancer (ESE) variants identified through the study of similar to 113,000 women of the BRIDGES cohort. A RAD51D minigene with exons 2-9 was constructed in splicing vector pSAD. Eleven BRIDGES splice-site variants (selected by MaxEntScan) were introduced into the minigene by site-directed mutagenesis and tested in MCF-7 cells. The 11 variants disrupted splicing, collectively generating 25 different aberrant transcripts. All variants but one produced negligible levels (<3.4%) of the full-length (FL) transcript. In addition, ESE elements of the alternative exon 3 were mapped by testing four overlapping exonic microdeletions (>= 30-bp), revealing an ESE-rich interval (c.202_235del) with critical sequences for exon 3 recognition that might have been affected by germline variants. Next, 26 BRIDGES variants and 16 artificial exon 3 single-nucleotide substitutions were also assayed. Thirty variants impaired splicing with variable amounts (0-65.1%) of the FL transcript, although only c.202G>A demonstrated a complete aberrant splicing pattern without the FL transcript. On the other hand, c.214T>C increased efficiency of exon 3 recognition, so only the FL transcript was detected (100%). In conclusion, 41 RAD51D spliceogenic variants (28 of which were from the BRIDGES cohort) were identified by minigene assays. We show that minigene-based mapping of ESEs is a powerful approach for identifying ESE hotspots and ESE-disrupting variants. Finally, we have classified nine variants as likely pathogenic according to ACMG/AMP-based guidelines, highlighting the complex relationship between splicing alterations and variant interpretation. | - |
dc.format.extent | 20 p. | - |
dc.format.mimetype | application/pdf | - |
dc.language.iso | eng | - |
dc.publisher | MDPI | - |
dc.relation.isformatof | Reproducció del document publicat a: https://doi.org/10.3390/cancers13112845 | - |
dc.relation.ispartof | Cancers, 2021, vol. 13, num. 11 | - |
dc.relation.uri | https://doi.org/10.3390/cancers13112845 | - |
dc.rights | cc by (c) Bueno Martínez et al., 2021 | - |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/es/ | * |
dc.source | Articles publicats en revistes (Institut d'lnvestigació Biomèdica de Bellvitge (IDIBELL)) | - |
dc.subject.classification | Càncer de mama | - |
dc.subject.classification | Càncer d'ovari | - |
dc.subject.other | Breast cancer | - |
dc.subject.other | Ovarian cancer | - |
dc.title | RAD51D Aberrant Splicing in Breast Cancer: Identification of Splicing Regulatory Elements and Minigene-Based Evaluation of 53 DNA Variants | - |
dc.type | info:eu-repo/semantics/article | - |
dc.type | info:eu-repo/semantics/publishedVersion | - |
dc.date.updated | 2021-06-25T09:14:44Z | - |
dc.relation.projectID | info:eu-repo/grantAgreement/EC/H2020/634935/EU//BRIDGES | - |
dc.rights.accessRights | info:eu-repo/semantics/openAccess | - |
Appears in Collections: | Publicacions de projectes de recerca finançats per la UE Articles publicats en revistes (Institut d'lnvestigació Biomèdica de Bellvitge (IDIBELL)) |
Files in This Item:
File | Description | Size | Format | |
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cancers-13-02845-v2.pdf | 1.81 MB | Adobe PDF | View/Open |
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