STRUCTURE OF THE PROMOTES AND COMPLETE SEQUENCE OF THE GENE CODING FOR THE RABBIT TRANSLATIONALLY CONTROLLED TUMOR PROTEIN (TCTP) P23

We have isolated genomic recombinants containing the complete gene cod ing for the rabbit translationally controlled tumor protein (TCTP), al so known as histamine-releasing factor (HRF) P23. The gene is organize d into five introns and six exons and its total length amounts to 3819 nucleotides. All intron/exon boundaries an in accordance with the GT/ AG rule. Transcription of the gene generates two mRNAs of 843 and 1163 nucleotides differing in the length of their 3'-untranslated regions. They are formed by alternative polyadenylation. The transcription ini tiation site has been determined by comparison of sequences of the gen e and several processed TCTP pseudogenes. The full-length 5'-untransla ted region comprises 116 nucleotides and starts with an oligopyrimidin e tract important for translational regulation. Additionally 1.2 kb of the 5'-flanking promoter region has been sequenced. The promoter cont ains a TATA box at -30 and potential binding sites for transcription f actors such as stimulating protein 1 (Sp1), nuclear factor 1 (NF1), ac tivator protein 1 (API), c-Ets1, cAMP-response element (CP2), myeloid- specific zinc finger protein 1 (MZF1) and others. For functional analy sis 5'-flanking sequences up to -918 were fused to the chloramphenicol acetyltransferase (CAT) gene and tested using a rabbit aortic smooch- muscle cell line by cell transfection and CAT assays. The results conf irm that the analyzed gene is the actively transcribed TCTP gene.