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Regulation of mRNA splicing and transport by the tyrosine kinase activity of src
1Institut de Recherches sur le Cancer, CNRS UPR 9044 Génétique Moléculaire et Intégration des Fonctions Cellulaires, Villejuif, France.
Abstract:
The regulation of transcription by signal transduction pathways is well documented. In addition, we have previously shown that src can regulate pre-mRNA processing. To investigate which functional domains of src are involved in the regulation of splicing and transport of Lymphotoxin alpha (LTalpha) transcripts, we have used src mutants in the catalytic, SH2 and SH3 domains in association with the Y527F or the E378G activating mutation. Our results establish that the regulation of pre-mRNA processing and transcription can occur independently of each other. The splicing and transport phenotypes require an intact tyrosine kinase domain and both are insensitive to the deletion of the SH3 domain. Therefore these phenotypes do not depend upon the recruitment through the SH3 domain of src of RNA binding proteins (Sam 68, hnRNP K). By contrast, deletions in the SH2 domain have no effect on splicing but either abolish or exacerbate the transport phenotype depending upon the activating mutation (Y527F or E378G). These divergent responses are associated with specific changes in the pattern of tyrosine phosphorylated proteins. Thus, the regulation of transcription, splicing and mRNA transport implicate different effector pathways of src. Furthermore, analysis of the transport phenotype reveals the interplay between the SH2 and catalytic domain of the protein.
Insights
Signal transduction pathways regulate transcription and pre-mRNA processing. Src
Area of Science:
- Molecular Biology
- Cell Signaling
- Gene Expression Regulation
Background:
- Signal transduction pathways are known regulators of transcription.
- Previous studies demonstrated src's role in pre-mRNA processing.
- Investigating src's functional domains in splicing and transport is crucial.
Purpose of the Study:
- To determine which src functional domains regulate Lymphotoxin alpha (LTalpha) transcript splicing and transport.
- To differentiate the roles of src's catalytic, SH2, and SH3 domains in these processes.
Main Methods:
- Utilized src mutants targeting catalytic, SH2, and SH3 domains.
- Employed Y527F or E378G activating mutations in src.
- Analyzed splicing and transport phenotypes of LTalpha transcripts.
Main Results:
- Pre-mRNA processing and transcription regulation occur independently.
- Splicing and transport require an intact tyrosine kinase domain, independent of the SH3 domain.
- SH2 domain deletions differentially affect transport based on activating mutations, correlating with tyrosine phosphorylation patterns.
- Src's SH2 and catalytic domains interact in regulating transport.
Conclusions:
- Src regulates transcription, splicing, and mRNA transport via distinct effector pathways.
- The SH3 domain is not essential for src-mediated splicing or transport.
- The SH2 domain plays a critical, mutation-dependent role in mRNA transport, highlighting domain interplay.