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Sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular RNAs
Conversely to canonical splicing, back-splicing connects the upstream 3' splice site (SS) with a downstream 5'SS and generates exonic circular RNAs (circRNAs) that are widely identified and have regulatory functions in eukaryotic gene expression. However, sex-specific back-splicing in Dros...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10250224/ https://www.ncbi.nlm.nih.gov/pubmed/37070178 http://dx.doi.org/10.1093/nar/gkad280 |
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author | Fan, Yu-Jie Ding, Zhan Zhang, Yu Su, Ruibao Yue, Jia-Le Liang, An-Min Huang, Qi-Wei Meng, Yan-Ran Li, Muwang Xue, Yuanchao Xu, Yong-Zhen |
author_facet | Fan, Yu-Jie Ding, Zhan Zhang, Yu Su, Ruibao Yue, Jia-Le Liang, An-Min Huang, Qi-Wei Meng, Yan-Ran Li, Muwang Xue, Yuanchao Xu, Yong-Zhen |
author_sort | Fan, Yu-Jie |
collection | PubMed |
description | Conversely to canonical splicing, back-splicing connects the upstream 3' splice site (SS) with a downstream 5'SS and generates exonic circular RNAs (circRNAs) that are widely identified and have regulatory functions in eukaryotic gene expression. However, sex-specific back-splicing in Drosophila has not been investigated and its regulation remains unclear. Here, we performed multiple RNA analyses of a variety sex-specific Drosophila samples and identified over ten thousand circular RNAs, in which hundreds are sex-differentially and -specifically back-spliced. Intriguingly, we found that expression of SXL, an RNA-binding protein encoded by Sex-lethal (Sxl), the master Drosophila sex-determination gene that is only spliced into functional proteins in females, promoted back-splicing of many female-differential circRNAs in the male S2 cells, whereas expression of a SXL mutant (SXL(RRM)) did not promote those events. Using a monoclonal antibody, we further obtained the transcriptome-wide RNA-binding sites of SXL through PAR-CLIP. After splicing assay of mini-genes with mutations in the SXL-binding sites, we revealed that SXL-binding on flanking exons and introns of pre-mRNAs facilitates back-splicing, whereas SXL-binding on the circRNA exons inhibits back-splicing. This study provides strong evidence that SXL has a regulatory role in back-splicing to generate sex-specific and -differential circRNAs, as well as in the initiation of sex-determination cascade through canonical forward-splicing. |
format | Online Article Text |
id | pubmed-10250224 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-102502242023-06-10 Sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular RNAs Fan, Yu-Jie Ding, Zhan Zhang, Yu Su, Ruibao Yue, Jia-Le Liang, An-Min Huang, Qi-Wei Meng, Yan-Ran Li, Muwang Xue, Yuanchao Xu, Yong-Zhen Nucleic Acids Res RNA and RNA-protein complexes Conversely to canonical splicing, back-splicing connects the upstream 3' splice site (SS) with a downstream 5'SS and generates exonic circular RNAs (circRNAs) that are widely identified and have regulatory functions in eukaryotic gene expression. However, sex-specific back-splicing in Drosophila has not been investigated and its regulation remains unclear. Here, we performed multiple RNA analyses of a variety sex-specific Drosophila samples and identified over ten thousand circular RNAs, in which hundreds are sex-differentially and -specifically back-spliced. Intriguingly, we found that expression of SXL, an RNA-binding protein encoded by Sex-lethal (Sxl), the master Drosophila sex-determination gene that is only spliced into functional proteins in females, promoted back-splicing of many female-differential circRNAs in the male S2 cells, whereas expression of a SXL mutant (SXL(RRM)) did not promote those events. Using a monoclonal antibody, we further obtained the transcriptome-wide RNA-binding sites of SXL through PAR-CLIP. After splicing assay of mini-genes with mutations in the SXL-binding sites, we revealed that SXL-binding on flanking exons and introns of pre-mRNAs facilitates back-splicing, whereas SXL-binding on the circRNA exons inhibits back-splicing. This study provides strong evidence that SXL has a regulatory role in back-splicing to generate sex-specific and -differential circRNAs, as well as in the initiation of sex-determination cascade through canonical forward-splicing. Oxford University Press 2023-04-18 /pmc/articles/PMC10250224/ /pubmed/37070178 http://dx.doi.org/10.1093/nar/gkad280 Text en © The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | RNA and RNA-protein complexes Fan, Yu-Jie Ding, Zhan Zhang, Yu Su, Ruibao Yue, Jia-Le Liang, An-Min Huang, Qi-Wei Meng, Yan-Ran Li, Muwang Xue, Yuanchao Xu, Yong-Zhen Sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular RNAs |
title | Sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular RNAs |
title_full | Sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular RNAs |
title_fullStr | Sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular RNAs |
title_full_unstemmed | Sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular RNAs |
title_short | Sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular RNAs |
title_sort | sex-lethal regulates back-splicing and generation of the sex-differentially expressed circular rnas |
topic | RNA and RNA-protein complexes |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10250224/ https://www.ncbi.nlm.nih.gov/pubmed/37070178 http://dx.doi.org/10.1093/nar/gkad280 |
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