The mRNAs encoding the two angiotensin-converting isozymes are transcribed from the same gene by a tissue-specific choice of alternative transcription initiation sites

Abstract

The two tissue-specific isozymes of angiotensin-converting enzyme are encoded by two mRNAs. The 5-kilobase pulmonary mRNA (mRNAP) and the 2.5-kilobase testicular mRNA (mRNAT) have identical sequences near their 3'-ends, whereas each has a unique sequence toward its 5'-end. Here we report that the two mRNAs originate from the same gene by initiation of transcription at two alternative sites. We have isolated a rabbit genomic clone which encodes these mRNAs. The sequence organization of the genome is such that the unique sequence of mRNAP precedes that of mRNAT, which in turn precedes the sequence common to both species. For generation of mRNAP, the mRNAT-specific sequence is eliminated as an intron from the primary transcript by splicing. Transcription of mRNAT, on the other hand, initiates within this intron at the middle of the gene. The exact initiation sites of mRNAT and mRNAP have been determined by DNA sequencing; DNA, oligonucleotide, and antisense RNA protection assays; and primer extension assays. Analyses of the sequence upstream of the transcription initiation sites revealed the presence of putative binding sites of several known transcription factors including SP-1, AP-2, and IID.