scholarly journals Regulation of SR protein phosphorylation and alternative splicing by modulating kinetic interactions of SRPK1 with molecular chaperones

2009 ◽  
Vol 23 (4) ◽  
pp. 482-495 ◽  
Author(s):  
X.-Y. Zhong ◽  
J.-H. Ding ◽  
J. A. Adams ◽  
G. Ghosh ◽  
X.-D. Fu
Keyword(s):  
Alternative Splicing ◽  
Protein Phosphorylation ◽  
Molecular Chaperones ◽  
Sr Protein

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.

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 Akt2 Regulation of Cdc2-Like Kinases (Clk/Sty), Serine/Arginine-Rich (SR) Protein Phosphorylation, and Insulin-Induced Alternative Splicing of PKCβII Messenger Ribonucleic Acid

Endocrinology ◽  
2008 ◽  
Vol 150 (5) ◽  
pp. 2087-2097 ◽  
Author(s):  
Kun Jiang ◽  
Niketa A. Patel ◽  
James E. Watson ◽  
Hercules Apostolatos ◽  
Eden Kleiman ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Protein Phosphorylation ◽  
Kinase Inhibitors ◽  
Sr Proteins ◽  
Null Mouse ◽  
Phosphatidylinositol 3 Kinase ◽  
Sr Protein ◽  
Messenger Ribonucleic Acid ◽  
Extracellular Signals ◽  
Mouse Tissues

Serine/arginine-rich (SR) proteins play essential roles in the constitutive and regulated splicing of precursor mRNAs. Phosphorylation of the arginine/serine dipeptide-rich (RS) domain by SR protein kinases such as Cdc2-like kinases (Clk/Sty) modulates their subcellular localization and activation. However, it remains unclear how these kinases and their target SR proteins are regulated by extracellular signals. Regulation of protein kinase C βII (PKCβII) pre-mRNA alternative splicing via exon inclusion by Akt2, a central kinase in insulin action, involves phosphorylation of SR proteins. Here we showed that Akt2, in response to insulin, resulted in phosphorylation of Clk/Sty, which then altered SR protein phosphorylation in concert with Akt2. Insulin-stimulated PKCβII pre-mRNA splicing was blocked by Clk/Sty and phosphatidylinositol-3-kinase inhibitors, and diabetic Akt2-null mouse tissues had impaired phospho-Clk/Sty, SR protein phosphorylation, and PKCβII expression. Furthermore, we observed that Akt2 phosphorylated several SR proteins distinct from Clk/Sty in response to insulin. Akt2-catalyzed phosphorylation of Clk/Sty and SR proteins revealed a role for both kinases in splicing regulation indicating dual functions for Akt2 in response to insulin in this pathway.

scholarly journals Central role of Akt in regulating Clk/Sty, serine/arginine‐rich (SR) protein phosphorylation and alternative splicing

2006 ◽  
Vol 20 (4) ◽  
Author(s):  
Denise R. Cooper ◽  
Karen D. Corbin ◽  
James E. Watson ◽  
Masatoshi Hagiwara ◽  
Niketa A. Patel ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Protein Phosphorylation ◽  
Sr Protein

scholarly journals Enhancer of rudimentary homologue interacts with scaffold attachment factor B at the nuclear matrix to regulate SR protein phosphorylation

FEBS Journal ◽  
2017 ◽  
Vol 284 (15) ◽  
pp. 2482-2500 ◽  
Author(s):  
Sotiria Drakouli ◽  
Aggeliki Lyberopoulou ◽  
Maria Papathanassiou ◽  
Ilias Mylonis ◽  
Eleni Georgatsou
Keyword(s):  
Protein Phosphorylation ◽  
Nuclear Matrix ◽  
Sr Protein ◽  
Factor B ◽  
Enhancer Of Rudimentary ◽  
Attachment Factor

scholarly journals Functional properties of p54, a novel SR protein active in constitutive and alternative splicing.

1996 ◽  
Vol 16 (10) ◽  
pp. 5400-5408 ◽  
Author(s):  
W J Zhang ◽  
J Y Wu
Keyword(s):  
Alternative Splicing ◽  
Splice Site ◽  
Sr Proteins ◽  
Dependent Manner ◽  
Sr Protein ◽  
Protein Protein Interaction ◽  
Small Nuclear Ribonucleoprotein ◽  
S100 Extract ◽  
U2 Auxiliary Factor ◽  
Auxiliary Factor

The p54 protein was previously identified by its reactivity with an autoantiserum. We report here that p54 is a new member of the SR family of splicing factors, as judged from its structural, antigenic, and functional characteristics. Consistent with its identification as an SR protein, p54 can function as a constitutive splicing factor in complementing splicing-deficient HeLa cell S100 extract. However, p54 also shows properties distinct from those of other SR family members, p54 can directly interact with the 65-kDa subunit of U2 auxiliary factor (U2AF65), a protein associated with the 3' splice site. In addition, p54 interacts with other SR proteins but does not interact with the U1 small nuclear ribonucleoprotein U1-70K or the 35-kDa subunit of U2 auxiliary factor (U2AF35). This protein-protein interaction profile is different from those of prototypical SR proteins SC35 and ASF/SF2, both of which interact with U1-70K and U2AF35 but not with U2AF65. p54 promotes the use of the distal 5' splice site in E1A pre-mRNA alternative splicing, while the same site is suppressed by ASF/SF2 and SC35. These findings and the differential tissue distribution of p54 suggest that this novel SR protein may participate in regulation of alternative splicing in a tissue- and substrate-dependent manner.

scholarly journals Lnc RNA ‐dependent nuclear stress bodies promote intron retention through SR protein phosphorylation

2019 ◽  
Vol 39 (3) ◽  
Author(s):  
Kensuke Ninomiya ◽  
Shungo Adachi ◽  
Tohru Natsume ◽  
Junichi Iwakiri ◽  
Goro Terai ◽  
...  
Keyword(s):  
Protein Phosphorylation ◽  
Intron Retention ◽  
Sr Protein

scholarly journals Arginine Methylation Controls the Subcellular Localization and Functions of the Oncoprotein Splicing Factor SF2/ASF

2010 ◽  
Vol 30 (11) ◽  
pp. 2762-2774 ◽  
Author(s):  
Rahul Sinha ◽  
Eric Allemand ◽  
Zuo Zhang ◽  
Rotem Karni ◽  
Michael P. Myers ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Subcellular Localization ◽  
Posttranslational Modifications ◽  
Target Genes ◽  
Positive Charge ◽  
Arginine Methylation ◽  
Sr Protein ◽  
Functional Studies ◽  
Eukaryotic Proteomes ◽  
Cellular Compartments

ABSTRACT Alternative splicing and posttranslational modifications (PTMs) are major sources of protein diversity in eukaryotic proteomes. The SR protein SF2/ASF is an oncoprotein that functions in pre-mRNA splicing, with additional roles in other posttranscriptional and translational events. Functional studies of SR protein PTMs have focused exclusively on the reversible phosphorylation of Ser residues in the C-terminal RS domain. We confirmed that human SF2/ASF is methylated at residues R93, R97, and R109, which were identified in a global proteomic analysis of Arg methylation, and further investigated whether these methylated residues regulate the properties of SF2/ASF. We show that the three arginines additively control the subcellular localization of SF2/ASF and that both the positive charge and the methylation state are important. Mutations that block methylation and remove the positive charge result in the cytoplasmic accumulation of SF2/ASF. The consequent decrease in nuclear SF2/ASF levels prevents it from modulating the alternative splicing of target genes, results in higher translation stimulation, and abrogates the enhancement of nonsense-mediated mRNA decay. This study addresses the mechanisms by which Arg methylation and the associated positive charge regulate the activities of SF2/ASF and emphasizes the significance of localization control for an oncoprotein with multiple functions in different cellular compartments.

scholarly journals Characterization and comparison of four serine- and arginine-rich (SR) protein kinases

1997 ◽  
Vol 326 (3) ◽  
pp. 693-700 ◽  
Author(s):  
Oliver NAYLER ◽  
Stefan STAMM ◽  
Axel ULLRICH
Keyword(s):  
Alternative Splicing ◽  
Catalytic Domain ◽  
Expression Patterns ◽  
Sr Proteins ◽  
Amino Acid Sequences ◽  
Sr Protein ◽  
High Sequence Identity ◽  
Mouse Tissues ◽  
Transformed Cell Lines

Phosphorylated serine- and arginine-rich (SR) proteins are components of the spliceosomal complex, and have been implicated in the control of alternative splicing. Kinases that regulate the phosphorylation and possibly the intranuclear distribution of SR proteins may therefore contribute to changes in choice of splice site. We have cloned three mouse cDNAs with high sequence identity to the family of LAMMER kinases (i.e. kinases carrying the conserved signature EHLAMMERILG in the catalytic domain). A comparison of their amino acid sequences revealed two related subfamilies with high evolutionary conservation. We have compared the expression patterns of these proteins in mouse tissues and transformed cell lines with that of a previously cloned family member (mCLK1/STY), and detected various transcripts for each gene. This underlines previous findings of alternative splicing of mclk1/STY. Our results suggest that the proportions of products for each gene are regulated independently. We further demonstrate that all variants encode autophosphorylating proteins that can phosphorylate several biochemically purified SR proteins in vitro, leading to hyperphosphorylation of at least one SR protein in vivo. The observed tissue distributions and substrate specificities suggest that these kinases may all be constituents of a network of regulatory mechanisms that enable SR proteins to control RNA splicing.

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