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Mol Cell Biol. Oct 1990; 10(10): 5215–5225.
PMCID: PMC361203
Translation to near the distal end of the penultimate exon is required for normal levels of spliced triosephosphate isomerase mRNA.
J Cheng, M Fogel-Petrovic, and L E Maquat
Department of Human Genetics, Roswell Park Cancer Institute, Buffalo, New York 14263.
The translation of human triosephosphate isomerase (TPI) mRNA normally terminates at codon 249 within exon 7, the final exon. Frameshift and nonsense mutations within the TPI gene that cause translation to terminate prematurely at or upstream of codon 189, within exon 6, result in a decreased level of TPI mRNA (I.O. Daar and L.E. Maquat, Mol. Cell. Biol. 8:802-813, 1988). For all mutations in this group, the decrease is to the same extent, i.e., to approximately 20% of the normal level. We show here that a second group of nonsense mutations that cause translation to terminate prematurely at or downstream of codon 208, in exon 6, did not affect TPI mRNA abundance. Deletion analysis demonstrated that the abundance of translationally active TPI mRNA is a function of both the distance and the polarity of the nonsense codon relative to the final intron in TPI pre-mRNA. Our results indicate that if translating ribosomes are unable to progress to at least a certain position within the penultimate exon relative to the final intron, then the level of the corresponding mRNA will be abnormally low. Studies inhibiting RNA synthesis with dactinomycin demonstrated that a block in translation does not affect the half-life of mature TPI mRNA. The simplest interpretation of our data is that the translation of TPI mRNA in the cytoplasm facilitates the splicing of TPI pre-mRNA or the transport of TPI mRNA across the nuclear envelope or both.
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