Comprehensive identification of abnormally spliced RNAs and elucidation of their significance

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In Y14(RNAi) and mag-1(RNAi) animals, unspliced tra-2 RNA is leaked to the cytoplasm and unusually spliced to tra-2s RNA by the cytoplasmic splicing enzyme IRE-1. The protein produced from tra-2s RNA acts dominant-negatively against normal TRA-2 protein because transgenic animals expressing TRA-2s protein showed germline masculinization like Y14(RNAi) animals. Interestingly, the leakage of unspliced RNA caused by the inhibition of Y14 or MAG-1 was also observed for almost all genes examined so far. Moreover, inhibition of eIF4AIII and SAP130 also caused leakage of unspliced RNA to the cytoplasm. Double RNAi analysis indicated that Y14 interacts with both SAP130 and two other splicing-related factors. These results suggest that the EJC core subunits associate with pre-mRNA and function as the gatekeeper to watch and block premature export of unspliced RNA, thereby contributing to mRNA quality control in C. elegans.