Isolation and characterization of nuclear proteins that bind to T cell receptor V beta decamer motif

TCR V beta promoter contains a highly conserved decamer homologous to cAMP response element (CRE). Recent studies have identified this CRE decamer as the dominant transcription-activating element within the TCR V beta promoter. We have isolated cDNA clones, TCR-ATF1 and TCR-ATF2, encoding DNA-binding proteins that recognize this CRE motif. The nucleotide sequence of TCR-ATF1 has not previously been reported, whereas that of TCR-ATF2 was homologous to CRE-BP1, ATF-2, and mXBP. Both TCR-ATF1 and TCR-ATF2 shared a conserved leucine zipper and DNA binding motif with other CRE-binding proteins. TCR-ATF1 and TCR-ATF2 were expressed in all cell lines examined and in mouse embryos as early as 12.5 days. Despite binding to the same CRE motif, TCR-ATF1 and TCR-ATF2 were different from CREB in the fine nucleotide specificity. TCR-ATF bound methylated CRE and CRE mutant M4 (4C----G) that were not recognized by CREB. Additionally, TCR-ATF1 weakly recognized two other single nucleotide mutants of V beta-CRE that were not bound by TCR-ATF2 and CREB. We have further demonstrated that TCR beta-chain expression was immediately activated by cAMP. Such induction is likely mediated through V beta-CRE sequence, because the inclusion of V beta-CRE in a vector with minimum promoter (pBLCAT2) conferred the cAMP inducibility of CAT activity.