Characterization of the regulation of CD46 RNA alternative splicing
Here we present a detailed analysis of the alternative splicing regulation of human CD46, which generates different isoforms with distinct functions. CD46 is a ubiquitous membrane protein that protects host cells from complement and plays other roles in immunity, autophagy, and cell adhesion. CD46 d...
| Main Authors: | , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
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2018
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| Online Access: | https://hdl.handle.net/10356/89443 http://hdl.handle.net/10220/46266 |
| _version_ | 1811697262648098816 |
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| author | Tang, Sze Jing Luo, Shufang Ly, Phuong Thao Goh, Eling Roca, Xavier Ho, Jessie Jia Xin |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Tang, Sze Jing Luo, Shufang Ly, Phuong Thao Goh, Eling Roca, Xavier Ho, Jessie Jia Xin |
| author_sort | Tang, Sze Jing |
| collection | NTU |
| description | Here we present a detailed analysis of the alternative splicing regulation of human CD46, which generates different isoforms with distinct functions. CD46 is a ubiquitous membrane protein that protects host cells from complement and plays other roles in immunity, autophagy, and cell adhesion. CD46 deficiency causes an autoimmune disorder, and this protein is also involved in pathogen infection and cancer. Before this study, the mechanisms of CD46 alternative splicing remained unexplored even though dysregulation of this process has been associated with autoimmune diseases. We proved that the 5′ splice sites of CD46 cassette exons 7 and 8 encoding extracellular domains are defined by noncanonical mechanisms of base pairing to U1 small nuclear RNA. Next we characterized the regulation of CD46 cassette exon 13, whose inclusion or skipping generates different cytoplasmic tails with distinct functions. Using splicing minigenes, we identified multiple exonic and intronic splicing enhancers and silencers that regulate exon 13 inclusion via trans-acting splicing factors like PTBP1 and TIAL1. Interestingly, a common splicing activator such as SRSF1 appears to repress CD46 exon 13 inclusion. We also report that expression of CD46 mRNA isoforms is further regulated by non-sense-mediated mRNA decay and transcription speed. Finally, we successfully manipulated CD46 exon 13 inclusion using antisense oligonucleotides, opening up opportunities for functional studies of the isoforms as well as for therapeutics for autoimmune diseases. This study provides insight into CD46 alternative splicing regulation with implications for its function in the immune system and for genetic disease. |
| first_indexed | 2024-10-01T07:52:28Z |
| format | Journal Article |
| id | ntu-10356/89443 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T07:52:28Z |
| publishDate | 2018 |
| record_format | dspace |
| spelling | ntu-10356/894432023-02-28T17:02:52Z Characterization of the regulation of CD46 RNA alternative splicing Tang, Sze Jing Luo, Shufang Ly, Phuong Thao Goh, Eling Roca, Xavier Ho, Jessie Jia Xin School of Biological Sciences Alternative Splicing Complement System DRNTU::Science::Biological sciences Here we present a detailed analysis of the alternative splicing regulation of human CD46, which generates different isoforms with distinct functions. CD46 is a ubiquitous membrane protein that protects host cells from complement and plays other roles in immunity, autophagy, and cell adhesion. CD46 deficiency causes an autoimmune disorder, and this protein is also involved in pathogen infection and cancer. Before this study, the mechanisms of CD46 alternative splicing remained unexplored even though dysregulation of this process has been associated with autoimmune diseases. We proved that the 5′ splice sites of CD46 cassette exons 7 and 8 encoding extracellular domains are defined by noncanonical mechanisms of base pairing to U1 small nuclear RNA. Next we characterized the regulation of CD46 cassette exon 13, whose inclusion or skipping generates different cytoplasmic tails with distinct functions. Using splicing minigenes, we identified multiple exonic and intronic splicing enhancers and silencers that regulate exon 13 inclusion via trans-acting splicing factors like PTBP1 and TIAL1. Interestingly, a common splicing activator such as SRSF1 appears to repress CD46 exon 13 inclusion. We also report that expression of CD46 mRNA isoforms is further regulated by non-sense-mediated mRNA decay and transcription speed. Finally, we successfully manipulated CD46 exon 13 inclusion using antisense oligonucleotides, opening up opportunities for functional studies of the isoforms as well as for therapeutics for autoimmune diseases. This study provides insight into CD46 alternative splicing regulation with implications for its function in the immune system and for genetic disease. MOE (Min. of Education, S’pore) Published version 2018-10-09T07:34:31Z 2019-12-06T17:25:35Z 2018-10-09T07:34:31Z 2019-12-06T17:25:35Z 2016 Journal Article Tang, S. J., Luo, S., Ho, J. J. X., Ly, P. T., Goh, E., & Roca, X. (2016). Characterization of the regulation of CD46 RNA alternative splicing. Journal of Biological Chemistry, 291(27), 14311-14323. doi:10.1074/jbc.M115.710350 0021-9258 https://hdl.handle.net/10356/89443 http://hdl.handle.net/10220/46266 10.1074/jbc.M115.710350 27226545 en Journal of Biological Chemistry © 2016 The American Society for Biochemistry and Molecular Biology, Inc. This paper was published in Journal of Biological Chemistry and is made available as an electronic reprint (preprint) with permission of The American Society for Biochemistry and Molecular Biology, Inc. The published version is available at: [http://dx.doi.org/10.1074/jbc.M115.710350]. One print or electronic copy may be made for personal use only. Systematic or multiple reproduction, distribution to multiple locations via electronic or other means, duplication of any material in this paper for a fee or for commercial purposes, or modification of the content of the paper is prohibited and is subject to penalties under law. 13 p. application/pdf |
| spellingShingle | Alternative Splicing Complement System DRNTU::Science::Biological sciences Tang, Sze Jing Luo, Shufang Ly, Phuong Thao Goh, Eling Roca, Xavier Ho, Jessie Jia Xin Characterization of the regulation of CD46 RNA alternative splicing |
| title | Characterization of the regulation of CD46 RNA alternative splicing |
| title_full | Characterization of the regulation of CD46 RNA alternative splicing |
| title_fullStr | Characterization of the regulation of CD46 RNA alternative splicing |
| title_full_unstemmed | Characterization of the regulation of CD46 RNA alternative splicing |
| title_short | Characterization of the regulation of CD46 RNA alternative splicing |
| title_sort | characterization of the regulation of cd46 rna alternative splicing |
| topic | Alternative Splicing Complement System DRNTU::Science::Biological sciences |
| url | https://hdl.handle.net/10356/89443 http://hdl.handle.net/10220/46266 |
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