Exonic splicing enhancers in fission yeast: functional conservation demonstrates an early evolutionary origin.
Discrete sequence elements known as exonic splicing enhancers (ESEs) have been shown to influence both the efficiency of splicing and the profile of mature mRNAs in multicellular eukaryotes. While the existence of ESEs has not been demonstrated previously in unicellular eukaryotes, the factors known to recognize these elements and mediate their communication with the core splicing machinery are conserved and essential in the fission yeast Schizosaccharomyces pombe. Here, we provide evidence that ESE function is conserved through evolution by demonstrating that three exonic splicing enhancers derived from vertebrates (chicken ASLV, mouse IgM, and human cTNT) promote splicing of two distinct S. pombe pre-messenger RNAs (pre-mRNAs). Second, as in extracts from mammalian cells, ESE function in S. pombe is compromised by mutations and increased distance from the 3'-splice site. Third, three-hybrid analyses indicate that the essential SR (serine/arginine-rich) protein Srp2p, but not the dispensable Srp1p, binds specifically to both native and heterologous purine-rich elements; thus, Srp2p is the likely mediator of ESE function in fission yeast. Finally, we have identified five natural purine-rich elements from S. pombe that promote splicing of our reporter pre-mRNAs. Taken together, these results provide strong evidence that the genesis of ESE-mediated splicing occurred early in eukaryotic evolution.
Webb
Christopher J
CJ
School of Medicine, Department of Molecular Biology and Microbiology, Case Western Reserve University, Cleveland, OH 44106-4960, USA.
Romfo
Charles M
CM
van Heeckeren
Willem J
WJ
Wise
Jo Ann
JA
D000818:Animals; D001358:Avian Sarcoma Viruses / Q000235:genetics; D002645:Chickens / Q000235:genetics; D019143:Evolution, Molecular; D006801:Humans; D007075:Immunoglobulin M / Q000235:genetics; D051379:Mice; D011485:Protein Binding / Q000235:genetics / Q000502:physiology; D012326:RNA Splicing / Q000235:genetics* / Q000502:physiology; D000072260:RNA Splicing Factors; D012333:RNA, Messenger / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D012568:Schizosaccharomyces / Q000235:genetics* / Q000502:physiology; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D007075:Immunoglobulin M; D000072260:RNA Splicing Factors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D029702:Schizosaccharomyces pombe Proteins; C110506:Srp1 protein, S pombe; C120856:srP2 protein, S pombe