Expression of human BRCA1Δ17–19 alternative splicing variant with a truncated BRCT domain in MCF-7 cells results in impaired assembly of DNA repair complexes and aberrant DNA damage response

Ševčı́k, Jan; Falk, Martin; Macůrek, Libor; Kleiblová, Petra; Lhota, Filip; Hojný, J.; Štefančíková, Lenka; Janatová, Markéta; Bártek, Jiří; Stříbrná, Jana; Hodný, Zdeněk; Ježková, Lucie; Pohlreich, Petr; Kleibl, Zdeněk

doi:10.1016/j.cellsig.2013.02.008

Cited by 25 publications

(29 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Several reports suggest that alternative splicing of specific genes, such as BRCA1, BRCA2, BARD1, or ERCC1, can impact on the response of cancer cells to PT and some studies also suggest that this correlates with PT resistance in human cancer [23][24][25][26][27][28]. Accordingly, it has been proposed that evaluation of alternative splicing of selected genes could serve as prognostic marker in ovarian cancer [29].…”

Section: Discussionmentioning

confidence: 99%

Splicing factor proline- and glutamine-rich (SFPQ) protein regulates platinum response in ovarian cancer-modulating SRSF2 activity

et al. 2020

View full text Add to dashboard Cite

In epithelial ovarian cancer (EOC), response to platinum (PT)-based chemotherapy dictates subsequent treatments and predicts patients' prognosis. Alternative splicing is often deregulated in human cancers and can be altered by chemotherapy. Whether and how changes in alternative splicing regulation could impact on the response of EOC to PT-based chemotherapy is still not clarified. We identified the splicing factor proline and glutamine rich (SFPQ) as a critical mediator of response to PT in an unbiased functional genomic screening in EOC cells and, using a large cohort of primary and recurrent EOC samples, we observed that it is frequently overexpressed in recurrent PT-treated samples and that its overexpression correlates with PT resistance. At mechanistic level, we show that, under PT treatment, SFPQ, in complex with p54 nrb , binds and regulates the activity of the splicing factor SRSF2. SFPQ/p54 nrb complex decreases SRSF2 binding to caspase-9 RNA, favoring the expression of its alternative spliced antiapoptotic form. As a consequence, SFPQ/p54 nrb protects cells from PTinduced death, eventually contributing to chemoresistance. Overall, our work unveils a previously unreported SFPQ/p54 nrb / SRSF2 pathway that in EOC cells plays a central role in regulating alternative splicing and PT-induced apoptosis and that could result in the design of new possible ways of intervention to overcome PT resistance.

show abstract

Section: Discussionmentioning

confidence: 99%

Splicing factor proline- and glutamine-rich (SFPQ) protein regulates platinum response in ovarian cancer-modulating SRSF2 activity

et al. 2020

View full text Add to dashboard Cite

show abstract

“…A quantitative analysis of the range of naturally occurring isoforms relative to full-length expression and relative to other BRCA1 or BRCA2 isoforms is required, as is a comprehensive analysis of the functional role of each of these isoforms in both the healthy functioning of BRCA genes and the consequences of sequence variation on this process (29). It will also be important to extend this investigation to breast and ovarian tissue, to gain a broader understanding of the tissue-specific nature of splice-isoform regulation.…”

Section: Discussionmentioning

confidence: 99%

Comparison of mRNA Splicing Assay Protocols across Multiple Laboratories: Recommendations for Best Practice in Standardized Clinical Testing

et al. 2014

View full text Add to dashboard Cite

Background Accurate evaluation of unclassified sequence variants in cancer predisposition genes is essential for clinical management and depends on a multifactorial analysis of clinical, genetic, pathologic, and bioinformatic variables and assays of transcript length and abundance. The integrity of assay data in turn relies on appropriate assay design, interpretation, and reporting. Methods We conducted a multicenter investigation to compare mRNA splicing assay protocols used by members of the ENIGMA (Evidence-Based Network for the Interpretation of Germline Mutant Alleles) consortium. We compared similarities and differences in results derived from analysis of a panel of breast cancer 1, early onset (BRCA1) and breast cancer 2, early onset (BRCA2) gene variants known to alter splicing (BRCA1: c.135-1G>T, c.591C>T, c.594-2A>C, c.671-2A>G, and c.5467+5G>C and BRCA2: c.426-12_8delGTTTT, c.7988A>T, c.8632+1G>A, and c.9501+3A>T). Differences in protocols were then assessed to determine which elements were critical in reliable assay design. Results PCR primer design strategies, PCR conditions, and product detection methods, combined with a prior knowledge of expected alternative transcripts, were the key factors for accurate splicing assay results. For example, because of the position of primers and PCR extension times, several isoforms associated with BRCA1, c.594-2A>C and c.671-2A>G, were not detected by many sites. Variation was most evident for the detection of low-abundance transcripts (e.g., BRCA2 c.8632+1G>A Δ19,20 and BRCA1 c.135-1g>t Δ5q and Δ3). Detection of low-abundance transcripts was sometimes addressed by using more analytically sensitive detection methods (e.g., BRCA2 c.426-12_8delGTTTT ins18bp). Conclusions We provide recommendations for best practice and raise key issues to consider when designing mRNA assays for evaluation of unclassified sequence variants.

show abstract

“…Textbook examples which support the importance of alternative splicing and its influence on protein functions are certain BRCA1 alternative splicing variants (ASVs), which lead to translation into protein isoforms lacking important conservative domains. As a result, proteins with a low functional level are formed and negatively influence/regulate the BRCA1 biological function 17,18 . The knowledge of the expression levels of HNF1B mRNA variants or protein isoforms is crucial for the precise interpretation of HNF1B as a prognostic marker in a wide spectrum of expression studies.…”

mentioning

confidence: 99%

Identification of novel HNF1B mRNA splicing variants and their qualitative and semi-quantitative profile in selected healthy and tumour tissues

Hojný

Bártů

Krkavcová

et al. 2020

Sci Rep

Self Cite

View full text Add to dashboard Cite

Hepatocyte nuclear factor-1-beta (HNF1B) is a transcription factor crucial for the development of several tissues, and a promising biomarker of certain solid tumours. Thus far, two HNF1B alternative splicing variants (ASVs) have been described, however, the complete spectrum, prevalence and role of HNF1B ASVs in tumorigenesis are unclear. Considering the equivocal data about HNF1B ASVs and expression presented in literature, our aim was to characterize the spectrum of HNF1B mRNA splicing variants across different tissues. Here, we characterize HNF1B ASVs with high sensitivity in carcinomas of the uterine corpus, large intestine, kidney, pancreas, and prostate, with selected paired healthy tissues, using the previously described multiplex PCR and NGS approach. We identified 45 ASVs, of which 43 were novel. The spectrum and relative quantity of expressed ASVs mRNA differed among the analysed tissue types. Two known (3p, Δ7_8) and two novel (Δ7, Δ8) ASVs with unknown biological functions were detected in all the analysed tissues in a higher proportion. Our study reveals the wide spectrum of HNF1B ASVs in selected tissues. Characterization of the HNF1B ASVs is an important prerequisite for further expression studies to delineate the HNF1B splicing pattern, potential ASVs functional impact, and eventual refinement of HNF1B's biomarker role. Hepatocyte nuclear factor 1 beta (HNF1B, also known as Transcription Factor-2, TCF2), belongs to a family of transcription factors which are crucial for the regulation of the development of various tissues and organs during embryogenesis. Although originally described in the liver, HNF1B also plays an important role in the development and differentiation of the kidney, pancreas, reproductive tract, and biliary system 1-3. The HNF1B gene comprises 9 exons and codes for a protein with 3 important functional domains: the N-terminal dimerization domain, the DNA-binding domain (consisting of the Pit1/Oct-1/Unc-86-POU-homeodomain and a POU-specific domain), and the C-terminal transactivation domain (Fig. 1) 4. Apart from its role during organogenesis in the embryonic stage, in adults HNF1B acts as a classic transcription activator of the expression of multiple genes implicated in cell cycle regulation, apoptosis, glucose metabolism 5-7 , and as a regulator of the expression of genes associated with stem or progenitor cells 3. HNF1B is expressed mainly in tubule-forming epithelial tissues, such as kidney or pancreatic exocrine duct tubules, and also in the gall bladder, colon, duodenum, intestine, lung, stomach, urinary bladder, liver, endometrium, prostate, testis, and appendix 3,8 .

show abstract

Expression of human BRCA1Δ17–19 alternative splicing variant with a truncated BRCT domain in MCF-7 cells results in impaired assembly of DNA repair complexes and aberrant DNA damage response

Cited by 25 publications

References 51 publications

Splicing factor proline- and glutamine-rich (SFPQ) protein regulates platinum response in ovarian cancer-modulating SRSF2 activity

Splicing factor proline- and glutamine-rich (SFPQ) protein regulates platinum response in ovarian cancer-modulating SRSF2 activity

Comparison of mRNA Splicing Assay Protocols across Multiple Laboratories: Recommendations for Best Practice in Standardized Clinical Testing

Identification of novel HNF1B mRNA splicing variants and their qualitative and semi-quantitative profile in selected healthy and tumour tissues

Contact Info

Product

Resources

About