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Experimental evidence for RNA trans‐splicing in mammalian cells.
Author(s) -
Eul J.,
Graessmann M.,
Graessmann A.
Publication year - 1995
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1995.tb07325.x
Subject(s) - biology , trans splicing , rna splicing , rna , genetics , microbiology and biotechnology , computational biology , gene
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.