Exon 9 skipping of apoptotic caspase-2 pre-mRNA is promoted by SRSF3 through interaction with exon 8
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Authors
Jang, Ha NaLee, Minho
Loh, Tiing Jen
Choi, Seung-Woo
Oh, Hyun Kyung
Moon, Heegyum
Cho, Sunghee
Hong, Seong-Eui
Kim, Do Han
Sheng, Zhi
Green, Michael R.
Park, Daeho
Zheng, Xuexiu
Shen, Haihong
Document Type
Journal ArticlePublication Date
2014-01-01Keywords
Alternative SplicingApoptosis
Binding Sites
Caspase 2
Exons
Gene Knockdown Techniques
HEK293 Cells
Humans
Mutagenesis
RNA Precursors
RNA-Binding Proteins
Sequence Deletion
Anti-apoptotic
Apoptotic
Caspase-2
Exon 9
Pre-mRNA splicing
SRSF3
Biochemistry
Genetics and Genomics
Molecular Biology
Metadata
Show full item recordAbstract
Alternative splicing plays an important role in gene expression by producing different proteins from a gene. Caspase-2 pre-mRNA produces anti-apoptotic Casp-2S and pro-apoptotic Casp-2L proteins through exon 9 inclusion or skipping. However, the molecular mechanisms of exon 9 splicing are not well understood. Here we show that knockdown of SRSF3 (also known as SRp20) with siRNA induced significant increase of endogenous exon 9 inclusion. In addition, overexpression of SRSF3 promoted exon 9 skipping. Thus we conclude that SRSF3 promotes exon 9 skipping. In order to understand the functional target of SRSF3 on caspase-2 pre-mRNA, we performed substitution and deletion mutagenesis on the potential SRSF3 binding sites that were predicted from previous reports. We demonstrate that substitution mutagenesis of the potential SRSF3 binding site on exon 8 severely disrupted the effects of SRSF3 on exon 9 skipping. Furthermore, with the approach of RNA pulldown and immunoblotting analysis we show that SRSF3 interacts with the potential SRSF3 binding RNA sequence on exon 8 but not with the mutant RNA sequence. In addition, we show that a deletion of 26nt RNA from 5' end of exon 8, a 33nt RNA from 3' end of exon 10 and a 2225nt RNA from intron 9 did not compromise the function of SRSF3 on exon 9 splicing. Therefore we conclude that SRSF3 promotes exon 9 skipping of caspase-2 pre-mRNA by interacting with exon 8. Our results reveal a novel mechanism of caspase-2 pre-mRNA splicing.Source
Jang HN, Lee M, Loh TJ, Choi SW, Oh HK, Moon H, Cho S, Hong SE, Kim do H, Sheng Z, Green MR, Park D, Zheng X, Shen H. Exon 9 skipping of apoptotic caspase-2 pre-mRNA is promoted by SRSF3 through interaction with exon 8. Biochim Biophys Acta. 2014 Jan;1839(1):25-32. doi: 10.1016/j.bbagrm.2013.11.006. Link to article on publisher's siteDOI
10.1016/j.bbagrm.2013.11.006Permanent Link to this Item
http://hdl.handle.net/20.500.14038/44027PubMed ID
24321384Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.1016/j.bbagrm.2013.11.006