Fas-activated serine/threonine kinase (FAST K) synergizes with TIA-1/TIAR proteins to regulate Fas alternative splicing.
The factors and mechanisms that mediate the effects of intracellular signaling cascades on alternative pre-mRNA splicing are poorly understood. TIA-1 (T-cell intracellular antigen 1) and TIAR (TIA-1-related) proteins regulate alternative pre-mRNA splicing by promoting the use of suboptimal 5' splice sites followed by uridine-rich intronic enhancer sequences. These proteins promote, for example, inclusion of Fas receptor exon 6, which leads to an mRNA encoding a pro-apoptotic form of the receptor at the expense of the form that skips exon 6, which encodes an anti-apoptotic form. Fas-activated serine/threonine kinase (FAST K) is known to interact with and phosphorylate TIA-1. Here we have tested the possibility that FAST K influences alternative pre-mRNA splicing by affecting the activity of TIA-1/TIAR. Depletion of FAST K form Jurkat cells leads to skipping of exon 6 from endogenous Fas transcripts. Conversely, FAST K overexpression enhances exon 6 inclusion of Fas reporters transfected in HeLa cells. Consistent with the possibility that the effects of FAST K are mediated by changes in the function of TIA-1/TIAR, the effects of FAST K overexpression (i) are largely suppressed by depletion of TIA-1 and TIAR and (ii) are significantly compromised by mutation of a TIA-1/TIAR-responsive enhancer present downstream of exon 6 5' splice site. Furthermore, in vitro phosphorylation of TIA-1 by FAST K results in enhanced U1 snRNP recruitment. Interestingly, this enhancement is not due to increased binding of TIA-1 to the pre-mRNA. Taken together, the results connect Fas signaling with the activity of splicing factors that modulate Fas alternative splicing, suggesting the existence of an autoregulatory loop that could serve to amplify Fas responses.
Izquierdo
José M
JM
Centro de Biología Molecular "Severo Ochoa"-Universidad Autónoma de Madrid (CBMSO-UAM), Cantoblanco 28049 Madrid, Spain. jmizquirdo@cbm.uam.es
Valcárcel
Juan
J
D017398:Alternative Splicing; D017209:Apoptosis; D002467:Cell Nucleus / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D009154:Mutation; D010766:Phosphorylation; D039102:Poly(A)-Binding Proteins / Q000737:chemistry* / Q000502:physiology*; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000502:physiology*; D012323:RNA Processing, Post-Transcriptional; D015398:Signal Transduction; D000074019:T-Cell Intracellular Antigen-1; D019014:fas Receptor / Q000378:metabolism*
D039102:Poly(A)-Binding Proteins; D000074019:T-Cell Intracellular Antigen-1; C071558:TIA1 protein, human; D019014:fas Receptor; C095367:FASTK protein, human; D017346:Protein Serine-Threonine Kinases