scholarly journals Alternative Splicing Switches Potassium Channel Sensitivity to Protein Phosphorylation

2001 ◽  
Vol 276 (11) ◽  
pp. 7717-7720 ◽  
Author(s):  
Lijun Tian ◽  
Rory R. Duncan ◽  
Martin S. L. Hammond ◽  
Lorraine S. Coghill ◽  
Hua Wen ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Potassium Channel ◽  
Protein Phosphorylation

Abstract 805: Differential Regulation Of Cytokine-induced Alternative Splicing Of The TF Gene By Serine/Arginine Rich Protein Kinases

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Ursula Rauch ◽  
Andreas Eisenreich ◽  
Wolfgang Poller ◽  
Heinz-Peter Schultheiss
Keyword(s):  
Alternative Splicing ◽  
Protein Phosphorylation ◽  
Mrna Expression ◽  
Tissue Factor ◽  
Splice Variant ◽  
Sr Protein ◽  
Phosphorylation State ◽  
Post Induction ◽  
Tnf Α ◽  
Tf Gene

Background: Higher eukaryotes control gene expression and increase protein diversity by alternative splicing of pre-mRNA. The Cdc2-like kinase (Clk) family, DNA topoisomerase I (DNA topo I) or Akt kinase are involved in splicing control by regulating the phosphorylation state of serine/arginine rich (SR) proteins. We recently showed that alternatively spliced human tissue factor (asHTF), a soluble isoform of tissue factor (TF), the primary initiator of coagulation, is expressed in HUVECs in response to inflammatory cytokines. This study investigated the role of Clks, DNA topo I and the PI3K-Pathway in regulation of TF-splicing in TNF-α induced HUVECs. Methods: HUVECs were incubated with inhibitors of Clks, DNA-topo I or PI3K and were then stimulated with TNF-α. The SR protein phosphorylation state was determined 2 min post induction. The full length (fl) TF and asHTF mRNA were assessed 60 min post induction by Real-Time PCR. Proteins were measured 5 and 8 hours after stimulation by Western blots and the cell thrombogenicity was analyzed via a chromogenic assay. Results: TNF-α inceased the mRNA expression of asHTF and flTF in HUVECs. The Clk-inhibitor completely inhibited the TNF-α induced expression of asHTF and reduced flTF by 30 %. Inhibition of DNA topo I increased asHTF expression and reduced the flTF expression. Inhibition of the PI3K/Akt-pathway had no effect on TF mRNA expression. Reduced Clk-inhibition the TF activity by 50 % whereas DNA topo I inhibition significantly decreased the procoagulant TF activity 8 hours post TNF-α induction. The Clk- and DNA-topo I-inhibitors altered the SR-protein phosphorylation pattern post TNF-α-induction. Additionally resulted inhibition of Clks in the generation of a third TF mRNA-splice variant, TF-A. Conclusion: Selective inhibition of Clks or DNA topo I leads to alterations of SR-protein phosphorylation and affects the differential expression of TF isoforms, thereby modulating the thrombogenicity of HUVECs. The inhibition of Clks contributes to the generation of a third TF splice variant. The inhibition of these kinases gives new insights into the regulation of the TF gene splicing process, which may result in new therapeutic strategies for modulating cellular thrombogenicity.

Multiple potassium–channel components are produced by alternative splicing at the Shaker locus in Drosophila

Nature ◽  
1988 ◽  
Vol 331 (6152) ◽  
pp. 137-142 ◽  
Author(s):  
Thomas L. Schwarz ◽  
Bruce L. Tempel ◽  
Diane M. Papazian ◽  
Yuh Nung Jan ◽  
Lily Y. Jan
Keyword(s):  
Alternative Splicing ◽  
Potassium Channel

scholarly journals Interactions between two fission yeast serine/arginine-rich proteins and their modulation by phosphorylation

2002 ◽  
Vol 368 (2) ◽  
pp. 527-534 ◽  
Author(s):  
Zhaohua TANG ◽  
Norbert F. KÄUFER ◽  
Ren-Jang LIN
Keyword(s):  
Alternative Splicing ◽  
Protein Phosphorylation ◽  
Fission Yeast ◽  
Protein Function ◽  
Sr Proteins ◽  
Specific Protein ◽  
Glutathione S Transferase ◽  
Sr Protein ◽  
Random Screen ◽  
Related Proteins

The unexpected low number of genes in the human genome has triggered increasing attention to alternative pre-mRNA splicing, and serine/arginine-rich (SR) proteins have been correlated with the complex alternative splicing that is a characteristic of metazoans. SR proteins interact with RNA and splicing protein factors, and they also undergo reversible phosphorylation, thereby regulating constitutive and alternative splicing in mammals and Drosophila. However, it is not clear whether the features of SR proteins and alternative splicing are present in simple and genetically tractable organisms, such as yeasts. In the present study, we show that the SR-like proteins Srp1 and Srp2, found in the fission yeast Schizosaccharomyces pombe, interact with each other and the interaction is modulated by protein phosphorylation. By using Srp1 as bait in a yeast two-hybrid analysis, we specifically isolated Srp2 from a random screen. This Srp interaction was confirmed by a glutathione-S-transferase pull-down assay. We also found that the Srp1—Srp2 complex was phosphorylated at a reduced efficiency by a fission yeast SR-specific kinase, Dis1-suppression kinase (Dsk1). Conversely, Dsk1-mediated phosphorylation inhibited the formation of the Srp complex. These findings offer the first example in fission yeast for interactions between SR-related proteins and the modulation of the interactions by specific protein phosphorylation, suggesting that a mammalian-like SR protein function may exist in fission yeast.

scholarly journals Regulation of Alternative Splicing by Reversible Protein Phosphorylation

2007 ◽  
Vol 283 (3) ◽  
pp. 1223-1227 ◽  
Author(s):  
Stefan Stamm
Keyword(s):  
Alternative Splicing ◽  
Protein Phosphorylation ◽  
Protein Phosphatase ◽  
Single Gene ◽  
Biological Properties ◽  
Regulatory Proteins ◽  
Protein Phosphatase 1 ◽  
Splice Sites ◽  
Reversible Protein Phosphorylation ◽  
Selection Of

The vast majority of human protein-coding genes are subject to alternative splicing, which allows the generation of more than one protein isoform from a single gene. Cells can change alternative splicing patterns in response to a signal, which creates protein variants with different biological properties. The selection of alternative splice sites is governed by the dynamic formation of protein complexes on the processed pre-mRNA. A unique set of these splicing regulatory proteins assembles on different pre-mRNAs, generating a “splicing” or “messenger ribonucleoprotein code” that determines exon recognition. By influencing protein/protein and protein/RNA interactions, reversible protein phosphorylation modulates the assembly of regulatory proteins on pre-mRNA and therefore contributes to the splicing code. Studies of the serine/arginine-rich protein class of regulators identified different kinases and protein phosphatase 1 as the molecules that control reversible phosphorylation, which controls not only splice site selection, but also the localization of serine/arginine-rich proteins and mRNA export. The involvement of protein phosphatase 1 explains why second messengers like cAMP and ceramide that control the activity of this phosphatase influence alternative splicing. The emerging mechanistic links between splicing regulatory proteins and known signal transduction pathways now allow in detail the understanding how cellular signals modulate gene expression by influencing alternative splicing. This knowledge can be applied to human diseases that are caused by the selection of wrong splice sites.

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