Using RT-PCR and rapid amplification of 5' cDNA ends (5' RACE), we have cloned three previously unrecognized endothelin B receptor (EDNRB) transcripts from a human melanoma cell line. Three distinct types of cDNAs (EDNRBDelta1, Delta2 and Delta3) were identified. EDNRBDelta1 starts upstream of the published transcription start site of hEDNRB without splicing, whereas, EDNRBDelta2 and EDNRBDelta3 are alternatively spliced. EDNRBDelta1 and EDNRBDelta2 share the same transcription initiation site and are 560bp upstream of the conventional hEDNRB, whereas that of EDNRBDelta3 is 939bp upstream from that described for the conventional hEDNRB. Interestingly, many transcription factor motifs are detectable in the upstream regions of these transcription initiation sites. The predicted amino acid sequences reveal that EDNRBDelta1 and EDNRBDelta2 produce the same protein as the conventional hEDNRB, but EDNRBDelta3 would give rise to additional in-frame 89- or 83-amino-acid residues at the N-terminus. EDNRBDelta3 generates the same amino acid sequence at the C terminus, but utilizes the polyadenylation signal within the open reading frame, resulting in a shorter 3'UTR. These transcripts are widely expressed in human tissues, but their expression patterns vary between different tissues.