Trans splicing of nematode pre-messenger RNA in vitro.
In nematodes, a fraction of mRNAs contains a common 22 nucleotide 5' terminal spliced leader (SL) sequence derived by trans splicing. Here, we show that a cell-free extract prepared from developing embryos of the parasitic nematode Ascaris lumbricoides catalyzes accurate and efficient SL addition to a synthetic pre-mRNA at an authentic trans splice acceptor site. SL addition occurs via a trans splicing reaction that proceeds through Y-branched intermediates. The branchpoint is located at either of two adenosine residues located 18 and 19 nucleotides upstream of the splice acceptor site.
Hannon
G J
GJ
Department of Molecular Biology and Microbiology, Case Western Reserve University, School of Medicine, Cleveland, Ohio 44106.
Maroney
P A
PA
Denker
J A
JA
Nilsen
T W
TW
D000818:Animals; D001200:Ascaris / Q000235:genetics*; D001483:Base Sequence; D004722:Endoribonucleases; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D015345:Oligonucleotide Probes; D012322:RNA Precursors / Q000235:genetics*; D012326:RNA Splicing; D012333:RNA, Messenger / Q000235:genetics*; D016914:Ribonuclease H; D006163:Ribonuclease T1
D015345:Oligonucleotide Probes; D012322:RNA Precursors; D012333:RNA, Messenger; D004722:Endoribonucleases; D016914:Ribonuclease H; D006163:Ribonuclease T1