EWS/FLI alters 5'-splice site selection.
The chimeric gene EWS/FLI is present in at least 85% of Ewing's sarcomas as a result of chromosomal translocations. The resulting fusion protein contains the N terminus of the RNA-binding protein EWS and the ETS DNA-binding domain of the transcription factor FLI-1. Although EWS/FLI binds DNA and activates transcription, both EWS and EWS/FLI also interact with SF1 and U1C, essential components of the splicing machinery. Therefore, we tested the ability of EWS and EWS/FLI to alter 5'-splice site selection using an E1A gene in vivo splicing assay. We found that EWS/FLI, but not EWS, interfered with heterogeneous nuclear ribonucleoprotein A1-dependent splice site selection of E1A. Mutational analysis of EWS/FLI revealed that the ability to affect pre-mRNA splicing coincided with transforming activity. Therefore, EWS/FLI has the ability to influence splicing as well as transcription.
Knoop
L L
LL
Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, Tennessee 38105, USA.
Baker
S J
SJ
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D034441:Heterogeneous-Nuclear Ribonucleoproteins; D006801:Humans; D051379:Mice; D015514:Oncogene Proteins, Fusion / Q000502:physiology*; D051789:Proto-Oncogene Protein c-fli-1; D012326:RNA Splicing; D034802:RNA-Binding Protein EWS; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012261:Ribonucleoproteins / Q000235:genetics; D014157:Transcription Factors / Q000502:physiology*; D014170:Transformation, Genetic
D017931:DNA Primers; C106031:EWS-FLI fusion protein; D000076245:Heterogeneous Nuclear Ribonucleoprotein A1; D034481:Heterogeneous-Nuclear Ribonucleoprotein Group A-B; D034441:Heterogeneous-Nuclear Ribonucleoproteins; C000619657:Hnrnpa1 protein, mouse; D015514:Oncogene Proteins, Fusion; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; D012261:Ribonucleoproteins; D014157:Transcription Factors; C000619656:hnRNPA1 protein, human