Splicing of mRNA mediated by tRNA sequences in mouse cells |
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Authors: | Michela Zamboni Daniela Scarabino Glauco P Tocchini-Valentini |
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Institution: | Istituto di Biologia Cellulare, Consiglio Nazionale delle Ricerche, 00015 Monterotondo, Rome, Italy |
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Abstract: | tRNA splicing is essential for the formation of tRNAs and therefore for gene expression. A circularly permuted sequence of an amber-suppressor pre-tRNA gene was inserted into the sequence encoding the mouse NEMO protein. We demonstrated that, in mouse cells, the hybrid pre-tRNA/pre-mRNAs can be spliced precisely at the sites of the pre-tRNA intron. This splicing reaction produces functional tRNAs that suppress amber codons as well as translatable mRNAs that sustain the NF-κB activation pathway. The RNA molecules extracted from mouse cells were amplified by RT-PCR, and their sequences were determined, confirming the identity of the splice junctions. We then applied the Archaea-express technology, in which an archaeal RNA endonuclease is expressed in mouse cells. We show that both the endogenous eukaryal endonuclease and the archaeal one cleave the hybrid pre-tRNA/pre-mRNAs in the same manner with an additive effect. |
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Keywords: | pre-tRNA splicing hybrid pre-tRNA/pre-mRNA archaeal tRNA-endonuclease NF-κ B signaling |
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