SRSF6 Regulates the Alternative Splicing of the Apoptotic Fas Gene by Targeting a Novel RNA Sequence
Alternative splicing (AS) is a procedure during gene expression that allows the production of multiple mRNAs from a single gene, leading to a larger number of proteins with various functions. The alternative splicing (AS) of Fas (Apo-1/CD95) pre-mRNA can generate membrane-bound or soluble isoforms w...
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MDPI AG
2022-04-01
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| Series: | Cancers |
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| Online Access: | https://www.mdpi.com/2072-6694/14/8/1990 |
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| author | Namjeong Choi Ha Na Jang Jagyeong Oh Jiyeon Ha Hyungbin Park Xuexiu Zheng Sunjae Lee Haihong Shen |
| author_facet | Namjeong Choi Ha Na Jang Jagyeong Oh Jiyeon Ha Hyungbin Park Xuexiu Zheng Sunjae Lee Haihong Shen |
| author_sort | Namjeong Choi |
| collection | DOAJ |
| description | Alternative splicing (AS) is a procedure during gene expression that allows the production of multiple mRNAs from a single gene, leading to a larger number of proteins with various functions. The alternative splicing (AS) of Fas (Apo-1/CD95) pre-mRNA can generate membrane-bound or soluble isoforms with pro-apoptotic and anti-apoptotic functions. SRSF6, a member of the Serine/Arginine-rich protein family, plays essential roles in both constitutive and alternative splicing. Here, we identified SRSF6 as an important regulatory protein in Fas AS. The cassette exon inclusion of Fas was decreased by SRSF6-targeting shRNA treatment, but increased by SRSF6 overexpression. The deletion and substitution mutagenesis of the Fas minigene demonstrated that the UGCCAA sequence in the cassette exon of the Fas gene causes the functional disruption of SRSF6, indicating that these sequences are essential for SRSF6 function in Fas splicing. In addition, biotin-labeled RNA-pulldown and immunoblotting analysis showed that SRSF6 interacted with these RNA sequences. Mutagenesis in the splice-site strength alteration demonstrated that the 5′ splice-site, but not the 3′ splice-site, was required for the SRSF6 regulation of Fas pre-mRNA. In addition, a large-scale RNA-seq analysis using GTEX and TCGA indicated that while SRSF6 expression was correlated with Fas expression in normal tissues, the correlation was disrupted in tumors. Furthermore, high SRSF6 expression was linked to the high expression of pro-apoptotic and immune activation genes. Therefore, we identified a novel RNA target with 5′ splice-site dependence of SRSF6 in Fas pre-mRNA splicing, and a correlation between SRSF6 and Fas expression. |
| first_indexed | 2024-03-09T11:03:16Z |
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| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-09T11:03:16Z |
| publishDate | 2022-04-01 |
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| series | Cancers |
| spelling | doaj.art-5ce864eb61824d7f9a29f085086fc0a22023-12-01T01:07:54ZengMDPI AGCancers2072-66942022-04-01148199010.3390/cancers14081990SRSF6 Regulates the Alternative Splicing of the Apoptotic Fas Gene by Targeting a Novel RNA SequenceNamjeong Choi0Ha Na Jang1Jagyeong Oh2Jiyeon Ha3Hyungbin Park4Xuexiu Zheng5Sunjae Lee6Haihong Shen7School of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 61005, KoreaSchool of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 61005, KoreaSchool of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 61005, KoreaSchool of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 61005, KoreaSchool of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 61005, KoreaSchool of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 61005, KoreaSchool of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 61005, KoreaSchool of Life Sciences, Gwangju Institute of Science and Technology, Gwangju 61005, KoreaAlternative splicing (AS) is a procedure during gene expression that allows the production of multiple mRNAs from a single gene, leading to a larger number of proteins with various functions. The alternative splicing (AS) of Fas (Apo-1/CD95) pre-mRNA can generate membrane-bound or soluble isoforms with pro-apoptotic and anti-apoptotic functions. SRSF6, a member of the Serine/Arginine-rich protein family, plays essential roles in both constitutive and alternative splicing. Here, we identified SRSF6 as an important regulatory protein in Fas AS. The cassette exon inclusion of Fas was decreased by SRSF6-targeting shRNA treatment, but increased by SRSF6 overexpression. The deletion and substitution mutagenesis of the Fas minigene demonstrated that the UGCCAA sequence in the cassette exon of the Fas gene causes the functional disruption of SRSF6, indicating that these sequences are essential for SRSF6 function in Fas splicing. In addition, biotin-labeled RNA-pulldown and immunoblotting analysis showed that SRSF6 interacted with these RNA sequences. Mutagenesis in the splice-site strength alteration demonstrated that the 5′ splice-site, but not the 3′ splice-site, was required for the SRSF6 regulation of Fas pre-mRNA. In addition, a large-scale RNA-seq analysis using GTEX and TCGA indicated that while SRSF6 expression was correlated with Fas expression in normal tissues, the correlation was disrupted in tumors. Furthermore, high SRSF6 expression was linked to the high expression of pro-apoptotic and immune activation genes. Therefore, we identified a novel RNA target with 5′ splice-site dependence of SRSF6 in Fas pre-mRNA splicing, and a correlation between SRSF6 and Fas expression.https://www.mdpi.com/2072-6694/14/8/1990alternative splicingSRSF6Fas |
| spellingShingle | Namjeong Choi Ha Na Jang Jagyeong Oh Jiyeon Ha Hyungbin Park Xuexiu Zheng Sunjae Lee Haihong Shen SRSF6 Regulates the Alternative Splicing of the Apoptotic Fas Gene by Targeting a Novel RNA Sequence Cancers alternative splicing SRSF6 Fas |
| title | SRSF6 Regulates the Alternative Splicing of the Apoptotic Fas Gene by Targeting a Novel RNA Sequence |
| title_full | SRSF6 Regulates the Alternative Splicing of the Apoptotic Fas Gene by Targeting a Novel RNA Sequence |
| title_fullStr | SRSF6 Regulates the Alternative Splicing of the Apoptotic Fas Gene by Targeting a Novel RNA Sequence |
| title_full_unstemmed | SRSF6 Regulates the Alternative Splicing of the Apoptotic Fas Gene by Targeting a Novel RNA Sequence |
| title_short | SRSF6 Regulates the Alternative Splicing of the Apoptotic Fas Gene by Targeting a Novel RNA Sequence |
| title_sort | srsf6 regulates the alternative splicing of the apoptotic fas gene by targeting a novel rna sequence |
| topic | alternative splicing SRSF6 Fas |
| url | https://www.mdpi.com/2072-6694/14/8/1990 |
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