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Genetic dissection of the BRCA2 promoter and transcriptional impact of DNA variants

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 Fraile Bethencourt, Eugenia & Valenzuela Palomo, Alberto  &  Díez Gómez, Beatriz &  Infante, Mar &  Durán, Mercedes & Marcos, Germán  &  Lastra, Enrique & Gómez Barrero, Susana  & Velasco, Eladio A.  (2018-05 ) .Genetic dissection of the BRCA2 promoter and transcriptional impact of DNA variants.

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 Fraile Bethencourt, Eugenia & Valenzuela Palomo, Alberto  &  Díez Gómez, Beatriz &  Infante, Mar &  Durán, Mercedes & Marcos, Germán  &  Lastra, Enrique & Gómez Barrero, Susana  & Velasco, Eladio A. . 2018-05 .Genetic dissection of the BRCA2 promoter and transcriptional impact of DNA variants.

https://hdl.handle.net/20.500.12080/39610
dc.contributor.author  Fraile Bethencourt, Eugenia
dc.contributor.author Valenzuela Palomo, Alberto 
dc.contributor.author  Díez Gómez, Beatriz
dc.contributor.author  Infante, Mar
dc.contributor.author  Durán, Mercedes
dc.contributor.author Marcos, Germán 
dc.contributor.author  Lastra, Enrique
dc.contributor.author Gómez Barrero, Susana 
dc.contributor.author Velasco, Eladio A. 
dc.date.accessioned 2024-02-08T16:30:38Z
dc.date.available 2024-02-08T16:30:38Z
dc.date.created 2018-05
dc.date.issued 2018-05
dc.identifier.uri https://hdl.handle.net/20.500.12080/39610
dc.description.abstract Purpose Promoter mutations may affect transcription and can be associated with human diseases. However, the promoters of the breast cancer (BC) genes are not regularly screened. Our goal was to investigate the BRCA2 promoter in order to study a possible correlation between impaired transcription and disease. Methods The proximal and core promoter of the BRCA2 gene was sequenced in 95 high-risk BC patients. A BRCA2-promoter insert [¿ 938 to +312 from the transcription start site (TSS)] was generated and cloned into the firefly luciferase vector pGL4.10. Promoter variants and deletions were introduced by site-directed mutagenesis and quantified by Dual-Luciferase assays and semi-quantitative RT-PCR. Results Three different variants were detected in high-risk BC patients: rs3092989, rs206118, and rs563971900. Functional mapping of 13 overlapping deletions revealed four down-regulating segments (TSS positions): ¿59_¿10del/µdel3 (16% of activity of the wild-type construct), ¿104_¿55del/µdel4 (62%), ¿239_¿190del/µdel7 (39%), ¿464_¿415/µdel12 (78%), suggesting the presence therein of putative transcriptional activator motifs. Additionally, six microdeletions rendered lucif erase overexpression: +32_+81del/µdel1 (356%), ¿14_+36del/µdel2 (180%), ¿194_¿145del/µdel6 (154%), ¿284_¿235del/ µdel8 (168%), ¿329_¿280del/µdel9 (111%), and ¿509_¿460del/µdel13 (139%), which is indicative of repressor elements. Functional assays of 15 promoter variants (including those detected in patients) showed that ten of them significantly altered expression with seven up-regulating (113¿163%) and three down-regulating (rs551887850_G, rs570548398_T, rs55880202_T; 72¿83%) SNPs. Eight of them were located in an ENCODE-DNase Hypersensitive Cluster (TSS ¿185 to +105) where most active transcriptional motifs are known to be placed. Conclusions BRCA2 expression is highly sensitive to promoter variations as most of them induced relevant changes. Moreo ver, we mapped critical regions of the BRCA2 promoter that may constitute potential targets for regulatory variants. Three SNPs moderately decreased luciferase activity, but confirmation of its potential pathogenicity requires further analysis. These data reinforce the need to screen the promoter regions of breast cancer genes with a view to discovering novel deleterious mutations. Keywords Breast cancer · Ovarian cancer · Susceptibility genes · BRCA2 · Transcription · Promoter · Regulatory mutations · Luciferase assays es_ES
dc.format application/pdf es_ES
dc.language eng es_ES
dc.rights CC-BY es_ES
dc.rights.uri http://creativecommons.org/licenses/by/4.0/deed.es es_ES
dc.title Genetic dissection of the BRCA2 promoter and transcriptional impact of DNA variants es_ES
dc.type info:eu-repo/semantics/article es_ES
dc.rights.accessrights info:eu-repo/semantics/restrictedAccess es_ES
dc.identifier.location N/A es_ES


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