Arginine/serine-rich domains of SR proteins can function as activators of pre-mRNA splicing.
Serine/arginine (SR)-rich splicing factors contain an RNA binding domain and an arginine/serine (RS)-rich domain required for protein-protein interactions. In addition to their roles in the basic splicing reaction, SR proteins function as components of splicing enhancer complexes. Here, we investigate the role of RS domains in splicing enhancer function. Hybrid proteins containing RS domains fused to the MS2 RNA binding protein were tested in vitro with RNA substrates bearing an MS2 recognition sequence. These hybrid proteins activated splicing in nuclear extracts, but not in S100 extracts lacking SR proteins. However, intact recombinant SR proteins could complement the activity of the hybrid proteins in S100 extracts. These data demonstrate that RS domains function as splicing activators and suggest that the general and enhancer-dependent functions of SR proteins can be uncoupled.
Graveley
B R
BR
Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.
Maniatis
T
T
D001120:Arginine; D002213:Capsid / Q000235:genetics / Q000378:metabolism; D036022:Capsid Proteins; D004742:Enhancer Elements, Genetic / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D029543:Nucleocytoplasmic Transport Proteins; D017434:Protein Structure, Tertiary; D012322:RNA Precursors / Q000378:metabolism*; D012326:RNA Splicing / Q000502:physiology*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics; D012261:Ribonucleoproteins; D012694:Serine; D000068103:Serine-Arginine Splicing Factors; D017381:Spliceosomes / Q000737:chemistry / Q000378:metabolism
D036022:Capsid Proteins; D009687:Nuclear Proteins; D029543:Nucleocytoplasmic Transport Proteins; D012322:RNA Precursors; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; C073288:SRSF2 protein, human; D000068103:Serine-Arginine Splicing Factors; D012694:Serine; D001120:Arginine