Pollard, A. J., Krainer, A. R., Robson, S. C., Europe-Finner, G. N. (May 2002) Alternative splicing of the adenylyl cyclase stimulatory G-protein G alpha(s) is regulated by SF2/ASF and heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1) and involves the use of an unusual TG 3'-splice Site. Journal of Biological Chemistry, 277 (18). pp. 15241-51. ISSN 0021-9258 (Print)0021-9258
Abstract
The factors involved in regulating alternative splicing of the human adenylyl cyclase stimulatory G-protein G alpha(s) in different cell types remain undefined. We have designed a G alpha(s) minigene that retains the signals required for G alpha(s) alternative splicing in vivo. Employing transient transfection of human myometrial smooth muscle cells and HeLa cells, as well as in vitro splicing assays, we have provided evidence that the antagonistic splicing factors SF2/ASF and hnRNPA1 act as potent regulators of G alpha(s) isoform expression in these cells. Both SF2/ASF and hnRNPA1 control the selection of competing 5'-splice sites and respectively promote inclusion or skipping of the small cassette-type exon 3 of G alpha(s) transcripts, resulting in the generation of G alpha(s)-long and G alpha(s)-short mRNA isoforms. We have also provided evidence that SF2/ASF and hnRNPA1 play a role in 3'-splice site selection involving the use of a non-canonical TG 3'-splice site preceding exon 4. Using a score-matrix analysis to identify putative exonic enhancer sequences (ESEs), we found multiple high score ESE motifs for SF2/ASF, SC35, and SRp40 in exon 3 of G alpha(s). These results suggest that tissue-specific expression of SF2/ASF and hnRNPA1 governs the expression of alternative isoforms of G alpha(s) in these different cells types.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Adenylate Cyclase/*metabolism *Alternative Splicing Base Sequence DNA Primers Female GTP-Binding Protein alpha Subunits, Gs/*genetics Gene Expression Regulation HeLa Cells *Heterogeneous-Nuclear Ribonucleoprotein Group A-B Heterogeneous-Nuclear Ribonucleoproteins Humans Molecular Sequence Data Muscle, Smooth/metabolism Myometrium/metabolism Nuclear Proteins/*metabolism Promoter Regions, Genetic Protein Isoforms/genetics RNA Splicing RNA, Messenger/genetics RNA-Binding Proteins Restriction Mapping Ribonucleoproteins/*metabolism Transcription, Genetic |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > Alternative Splicing bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > G protein bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > nuclear ribonucleoprotein bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > splice site |
| CSHL Authors: | |
| Communities: | CSHL labs > Krainer lab |
| Depositing User: | Matt Covey |
| Date: | 3 May 2002 |
| Date Deposited: | 11 Mar 2014 20:24 |
| Last Modified: | 11 Mar 2014 20:24 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/29601 |
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