Dog CREM transcription factors: cloning, tissue distribution, and identification of new isoforms

CREM (cAMP Response Element Modulator) transcription factors are involved in the cAMP-dependent transcriptional regulation of CRE-containing genes. Multiple CREM transactivators and repressors are generated from a single gene by alternative splicings and use of an alternative intronic promoter. Here we report the cloning and sequencing of the full-length dog CREM cDNA, corresponding to the CREMtau alpha splice variant. Amino acid sequence identity with mouse and human orthologs reached 94.5% and 91.0% respectively. Using the RNAse Protection Assay (RPA) method with three distinct probes, we analyzed the expression of the various CREM transcripts in several dog tissues. We showed that CREM transcription factors have a restricted tissue distribution and that the ratio between activators and repressors varies considerably from one tissue to another. Moreover, we amplified, by RT-PCR, a cDNA that corresponds to two new CREM isoforms and confirmed, by RPA experiments, the presence of these mRNAs in dog thyroid and in other tissues. These transcripts result from splicing of the gamma domain and encode potential CREM transactivators (CREMtau alphagamma and CREMtau2 alphagamma).