SF2/ASF regulates proteomic diversity by affecting the balance between translation initiation mechanisms.
Post-splicing activities have been described for a subset of shuttling serine/arginine-rich splicing regulatory proteins, among them SF2/ASF. We showed that growth factors activate a Ras-PI 3-kinase-Akt/PKB signaling pathway that not only modifies alternative splicing of the fibronectin EDA exon, but also alters in vivo translation of reporter mRNAs containing the EDA binding motif for SF2/ASF, providing two co-regulated levels of isoform-specific amplification. Translation of most eukaryotic mRNAs is initiated via the scanning mechanism, which implicates recognition of the m7G cap at the mRNA 5'-terminus by the eIF4F protein complex. Several viral and cellular mRNAs are translated in a cap-independent manner by the action of cis-acting mRNA elements named internal ribosome entry sites that direct internal ribosome binding to the mRNA. Here we use bicistronic reporters that generate mRNAs carrying two open reading frames, one translated in a cap-dependent manner while the other by internal ribosome entry site-dependent initiation, to show that in vivo over-expression of SF2/ASF increases the ratio between cap-dependent and internal ribosome entry site-dependent translation. Consistently, knocking-down of SF2/ASF causes the opposite effect. Changes in expression levels of SF2/ASF also affect alternative translation of an endogenous mRNA, that one coding for fibroblast growth factor-2. These results strongly suggest a role for SF2/ASF as a regulator of alternative translation, meaning the generation of different proteins by the balance among these two translation initiation mechanisms, and expand the known potential of SF2/ASF to regulate proteomic diversity to the translation field.
Instituto de Fisiología, Biología Molecular y Neurociencias, Buenos Aires, Argentina.
Matías
M
Blaustein
Leandro
L
Quadrana
Guillermo
G
Risso
Manuel de la
Mde L
Mata
Federico
F
Pelisch
Anabella
A
Srebrow
D017398:Alternative Splicing; D002460:Cell Line; D016222:Fibroblast Growth Factor 2 / Q000235:genetics; D006801:Humans; D009687:Nuclear Proteins / Q000502:physiology*; D016366:Open Reading Frames; D010442:Peptide Chain Initiation, Translational; D014176:Protein Biosynthesis / Q000235:genetics*; D020543:Proteome / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000494:pharmacology; D016601:RNA-Binding Proteins; D012270:Ribosomes / Q000378:metabolism; D000068103:Serine-Arginine Splicing Factors
D009687:Nuclear Proteins; D020543:Proteome; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D016601:RNA-Binding Proteins; D016222:Fibroblast Growth Factor 2; D000068103:Serine-Arginine Splicing Factors