Functions of SR proteins in the U12-dependent AT-AC pre-mRNA splicing pathway.
SR proteins play critical roles in the major pre-mRNA splicing pathway. A second pathway processes U12-dependent AT-AC introns. We demonstrate, by biochemical complementation, the requirement for SR proteins in splicing of AT-AC introns. Whereas SR proteins were sufficient to activate splicing of a P120 AT-AC intron, splicing of a sodium channel AT-AC intron required an additional nuclear fraction. Individual recombinant SR proteins promoted splicing of both substrates, but displayed marked preferences. SR proteins supported basal AT-AC splicing, and also splicing stimulation via a downstream enhancer or conventional 5' splice site. Analysis of chimeric transcripts revealed that information dispersed throughout exons and introns dictates SR protein specificity and the requirement for the additional nuclear fraction. Thus, SR proteins function in both major and minor splicing pathways, and in coordinating the activities of both spliceosomes via exon definition. These results suggest that despite the substantial differences in intron consensus sequences and in four of the five snRNPs in each spliceosome, at least some of the interactions involving SR proteins are conserved between the two pathways.
Hastings
M L
ML
Cold Spring Harbor Laboratory, New York 11724-2208, USA.
Krainer
A R
AR
D002474:Cell-Free System; D005091:Exons; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D062553:NAV1.4 Voltage-Gated Sodium Channel; D009687:Nuclear Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D012322:RNA Precursors / Q000378:metabolism*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000378:metabolism*; D016601:RNA-Binding Proteins; D017411:Ribonucleoproteins, Small Nuclear / Q000378:metabolism*; D000068103:Serine-Arginine Splicing Factors; D015222:Sodium Channels / Q000235:genetics
D062553:NAV1.4 Voltage-Gated Sodium Channel; D009687:Nuclear Proteins; D010750:Phosphoproteins; D012322:RNA Precursors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D017411:Ribonucleoproteins, Small Nuclear; C106251:SCN4A protein, human; D015222:Sodium Channels; C552126:U11-U12 small nuclear ribonucleoprotein; D000068103:Serine-Arginine Splicing Factors