Regulation of Translational Efficiency by Disparate 5' UTRs of PPARgamma Splice Variants

The PPAR-gamma gene encodes for at least 7 unique transcripts due to alternative splicing of five exons in the 5'-untranslated region (UTR). The translated region is encoded by exons 1-6, which are identical in all isoforms. This study investigated the role of the 5'-UTR in regulating the efficiency with which the message is translated to protein. A coupled in vitro transcription-translation assay demonstrated that PPAR-gamma1, -gamma2, and -gamma5 are efficiently translated, whereas PPAR-gamma4 and -gamma7 are poorly translated. An in vivo reporter gene assay using each 5'-UTR upstream of the firefly luciferase gene showed that the 5'-UTRs for PPAR-gamma1, -gamma2, and -gamma4 enhanced translation, whereas the 5'-UTRs for PPAR-gamma5 and -gamma7 inhibited translation. Models of RNA secondary structure, obtained by the mfold software, were used to explain the mechanism of regulation by each 5'-UTR. In general, it was found that the translational efficiency was inversely correlated with the stability of the mRNA secondary structure, the presence of base-pairing in the consensus Kozak sequence, the number of start codons in the 5'-UTR, and the length of the 5'-UTR. A better understanding of posttranscriptional regulation of translation will allow modulation of protein levels without altering transcription.