Alternative pre-mRNA splicing is one of main contributors to protein diversity. A fundamental unanswered question is how splicing decision is altered by signal transduction. The C1 exon of the glutamate NMDA receptor NR1 subunit is alternatively spliced in a tissue-specific manner. It is predominantly included in the forebrain, and predominantly skipped in the hindbrain. Importantly, the C1 exon encodes the C1 protein cassette containing an ER retention signal that regulates the surface delivery rate of assembled receptors, and a calmodulin binding site that plays critical roles in learning and memory via regulating Ca2+ flux. Recently, it has been reported C1 exon skipping can be induced by cell depolarization. However, the molecular links mediating the depolarization on the cell membrane and alternative splicing in the nucleus largely remain unclear. Here I have investigated the roles of Ca2+ signaling, PKA and hnRNP A1 in mediating depolarization induced C1 splicing silencing. Specifically, using Ca2+ imaging, I have found the KCl induced sustained Ca2+ influx is associated with C1 splicing silencing. In addition, I have demonstrated that CaMK IV and PKA regulate C1 splicing in a hnRNP A1 dependent manner. Finally, I have shown that hnRNP A1 is recruited to the C1 splicing site upon depolarization in neurons. The results here have demonstrated signal transduction from depolarization to splicing factors, showing how splicing decision can be altered by external stimuli.
| Identifer | oai:union.ndltd.org:PITT/oai:PITTETD:etd-08182011-103915 |
| Date | 30 September 2011 |
| Creators | Zhang, Ping |
| Contributors | Roger Hendrix, Jeffrey L. Brodsky, Thomas E Smithgall, Paula Grabowski, Jeffrey Hildebrand |
| Publisher | University of Pittsburgh |
| Source Sets | University of Pittsburgh |
| Language | English |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.library.pitt.edu/ETD/available/etd-08182011-103915/ |
| Rights | unrestricted, I hereby certify that, if appropriate, I have obtained and attached hereto a written permission statement from the owner(s) of each third party copyrighted matter to be included in my thesis, dissertation, or project report, allowing distribution as specified below. I certify that the version I submitted is the same as that approved by my advisory committee. I hereby grant to University of Pittsburgh or its agents the non-exclusive license to archive and make accessible, under the conditions specified below, my thesis, dissertation, or project report in whole or in part in all forms of media, now or hereafter known. I retain all other ownership rights to the copyright of the thesis, dissertation or project report. I also retain the right to use in future works (such as articles or books) all or part of this thesis, dissertation, or project report. |
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