An in vitro system for the editing of apolipoprotein B mRNA.
A novel form of RNA editing generates two forms of apolipoprotein B (apo-B) mRNA by converting C at nucleotide 6666 to U or a U-like base. We have established an in vitro system for the editing of apo-B mRNA using synthetic RNAs and S100 extracts from rat hepatoma cells. Editing was detected by a sensitive primer extension assay and confirmed by DNA sequencing. The in vitro editing activity is specific and sensitive to proteinase K. Apo-B100 RNAs were synthesized in vitro from deletion mutants spanning nucleotide 6666. Synthetic RNAs containing 2383, 483, and 55 nucleotides of apo-B mRNA sequence were edited in vitro with similar efficiency, but an RNA containing 26 nucleotides was not edited.
Division of Molecular Medicine, MRC Clinical Research Centre, Harrow, Middlesex, England.
D M
DM
Driscoll
J K
JK
Wynne
S C
SC
Wallis
J
J
Scott
D000818:Animals; D001055:Apolipoproteins B / Q000235:genetics*; D001483:Base Sequence; D003001:Cloning, Molecular; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D008114:Liver Neoplasms, Experimental; D015345:Oligonucleotide Probes; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000235:genetics*; D051381:Rats; D013045:Species Specificity; D013329:Structure-Activity Relationship
D001055:Apolipoproteins B; D015345:Oligonucleotide Probes; D012333:RNA, Messenger