The 5′ untranslated regions of acetyl‐coenzyme A carboxylase mRNA provide specific translational control in vitro

Abstract

Acetyl-coenzyme A carboxylase (ACC) catalyzes the rate-limiting step in the biosynthesis of long-chain fatty acids. Transcription of the single-copy ACC gene from two independent promoters, together with the differential splicing of the transcripts, gives rise to mature ACC mRNA having the same open reading frame (ORF), but exhibiting heterogeneity in their 5' untranslated region (5'-UTR). Class 1 ACC mRNA are transcribed from the inducible promoter 1 and their 5'-end leading sequences are provided by exon 1. Class 2 ACC mRNA are transcribed from the constitutively expressing promoter 2 and their leading sequences are derived from exon 2. In order to understand the role of different 5' UTR of ACC mRNA we have synthesized in vitro transcripts with defined ACC mRNA 5' UTR and examined their relative translational efficiencies in rabbit reticulocyte lysates. The major translation product of both forms of ACC mRNA was initiated at the first AUG of the ORF. Class 1 transcripts had a 6-9-fold better translational efficiency than class 2 transcripts, based on the quantity of major peptide produced by a given amount of transcript. The poor translational efficiency of class 2 transcripts can be improved by the removal of sequences contributed by exon 2, suggesting that they play an inhibitory role in the translation of class 2 types of ACC mRNA. In addition to their higher translational efficiency, the class 1 transcripts can also initiate translation at in-frame non-AUG codons, located in exon 1, i.e. upstream to the starting AUG of the common ACC mRNA ORF. This results in novel ACC peptides with extended N termini. These observations are consistent with the hypothesis that the 5' UTR heterogeneity in the ACC mRNA may be involved in post-transcriptional control, at the level of translation, of the ACC gene expression.