Nucleotide sequence analysis of six independently isolated cDNAs for rat acyl-CoA synthetase (ACS) revealed three forms of ACS mRNA, designated form-A, -B, and -C mRNAs, which differ in their 5′-untranslated regions. Form-A mRNA was preferentially detected in normal and peroxisome-induced livers, whereas form-B mRNA was found in peroxisome-induced livers but not in normal livers and hearts, and form-C mRNA was preferentially found in normal hearts and peroxisome-induced livers. Analysis of two overlapping genomic clones for the rat ACS gene revealed that the three 5′-untranslated regions of the mRNAs are individually encoded by three different exons located within a 20-kilobase genomic fragment. The transcription start sites of the three forms of ACS mRNA were determined and nucleotide sequences of 5′-upstream regions of the three 5′-end exons were determined. The 5′-upstream regions were fused to the chloramphenicol acetyltransferase gene and transcription units of the three forms of ACS mRNAs were determined. These data indicate that the three forms of ACS mRNA with 5′-end heterogeneity are generated by alternative transcription from three promoters in the rat ACS gene.
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