Spurious splicing within the XIAP 5' UTR occurs in the Rluc/Fluc but not the betagal/CAT bicistronic reporter system.
X-chromosome-linked inhibitor of apoptosis, XIAP, has been shown to contain a strong internal ribosome entry site (IRES) within its 5' untranslated region (UTR) that promotes translation of XIAP mRNA under conditions of cellular stress. This claim came under scrutiny in a recent report demonstrating that the XIAP 5' UTR undergoes splicing when inserted between the two reporter cistrons of the dual luciferase plasmid Rluc/Fluc. In this paper, we demonstrate that the splicing within the XIAP 5' UTR specifically occurs only in the context of mRNA produced from the Rluc/Fluc but not the pbetagal/CAT bicistronic reporter plasmid.
Holcik
Martin
M
Apoptosis Research Center, Children's Hospital of Eastern Ontario, Ottawa, Canada. martin@arc.cheo.ca
Graber
Tyson
T
Lewis
Stephen M
SM
Lefebvre
Charles A
CA
Lacasse
Eric
E
Baird
Stephen
S
D020121:5' Untranslated Regions / Q000235:genetics* / Q000378:metabolism; D017398:Alternative Splicing; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D007668:Kidney / Q000378:metabolism; D049410:Luciferases, Renilla / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D012270:Ribosomes / Q000235:genetics; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D020121:5' Untranslated Regions; D012333:RNA, Messenger; D051636:X-Linked Inhibitor of Apoptosis Protein; D049410:Luciferases, Renilla; D015500:Chloramphenicol O-Acetyltransferase; D001616:beta-Galactosidase