Experimental evidence for RNA trans-splicing in mammalian cells.
We present evidence that mammalian cells have the ability to generate functional mRNA molecules by trans-splicing. Rat cells, transformed by an early SV40 DNA fragment (Bst/Bam) synthesize a truncated T antigen (T1 antigen), although the cells do not have a direct sequence homology for the T1 antigen at the DNA level. The Bst/Bam DNA fragment encodes exclusively for the second SV40 T antigen exon (aa 83-708) and contains the entire small t antigen intron. To synthesize the corresponding mRNA (T1 mRNA), the cells utilize a cryptic 5' splice site within the second exon (codons for aa 131/132) as donor site and the upstream small t antigen 3' splice site as the acceptor site. Since these sites are in an inverted order on the pre-mRNA, two Bst/Bam transcripts are required to generate one T1 mRNA molecule. HeLa cell nuclear extracts also performed the trans-splicing reaction in vitro.
Eul
J
J
Institut für Molekularbiologie und Biochemie, Freie Universität Berlin, Germany.
Graessmann
M
M
Graessmann
A
A
D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D000957:Antigens, Viral, Tumor / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D002478:Cells, Cultured; D005091:Exons; D015967:Gene Expression Regulation, Viral; D006367:HeLa Cells; D006801:Humans; D007438:Introns; D008322:Mammals / Q000235:genetics*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D012313:RNA / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics; D051381:Rats
D000957:Antigens, Viral, Tumor; D012333:RNA, Messenger; D012367:RNA, Viral; D012313:RNA