期刊
BMB REPORTS
卷 54, 期 3, 页码 176-181出版社
KOREAN SOCIETY BIOCHEMISTRY & MOLECULAR BIOLOGY
DOI: 10.5483/BMBRep.2021.54.3.170
关键词
5 ' splice-site; Alternative splicing; Bcl-x; SRSF2; SRSF6
资金
- Cell Logistics Research Center - Ministry of Education [2016R1A5A1007318]
- National Research Foundation of Korea
- GIST Research Institute (GRI) ARI grant - GIST in 2020
- [NRF-2020R1A2C2004682]
- [NRF-2019R1I 1A1A01057372]
The study identified SRSF2 and SRSF6 as regulatory factors for 5' splice-site selection of Bcl-x pre-mRNA, showing that a single binding motif or cluster is not their functional target. Mutagenesis of the 5' splice-site to a conserved or weaker score can limit the role of SRSF2 and SRSF6 in 5' splice-site activation.
Bcl-x, a member of the Bcl-2 family, plays a key role in apoptosis. Alternative splicing of Bcl-x pre-mRNA through alternative 5' splice-site selection produces an anti-apoptotic mRNA isoform that includes exon 2b and a pro-apoptotic Bcl-x mRNA isoform that excludes exon 2b. Here we used Bcl-x minigene and identified SRSF2 and SRSF6 as two regulatory factors of 5' splice-site selection of Bcl-x pre-mRNA. We selected binding clusters closer to 5' splice-sites from multiple potential binding sites of SRSF2 and SRSF6 to perform loss of functions analysis through site-directed mutagenesis. Our results demonstrated that these mutations did not abolish regulatory functions of SRSF2 or SRSF6, indicating that a single binding motif or a cluster was not a functional target of these proteins in Bcl-x pre-mRNA splicing. Random deletion mutagenesis did not disrupt the role of SRSF2 and SRSF6. Importantly, mutagenesis of 5' splice-site to a conserved or a weaker score demonstrated that the weaker strength of the target 5' splice-site or higher strength of the other 5' splice-site strength limited the role of SRSF2 and SRSF6 in 5' splice-site activation.
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