DIFFERENTIAL EXPRESSION OF RNA TRANSCRIPTS ENCODING UNIQUE CARBOXY-TERMINAL SEQUENCES OF HUMAN PARATHYROID HORMONE-RELATED PEPTIDE

The human gene encoding PTH-related peptide (hPTHrP) is a complex transcriptional unit that gives rise to multiple messenger RNA (mRNA) transcripts in normal and neoplastic tissues. The utilization of three different transcription start sites and alternative splicing of exons encoding 5'-untranslated sequences accounts for some of the heterogeneity in hPTHrP gene expression; however, the pattern and potential significance of alternative exon splicing at the 3'-end of the hPTHrP gene have not been systematically examined. We have used exon-specific primers and reverse transciptase-polymerase chain reaction to analyze hPTHrP gene expression in nonneoplastic human tissues and human tumors. PTHrP mRNA transcripts encoding PTHrP-(1-139), -(1-173), or -(1-141) were identified in the majority of tissues analyzed. PTHrP mRNAs containing either exon 6 [PTHrP-(1-139)] or exon 8 [PTHrP-(1-141)] sequences were considerably more abundant than mRNAs encoding PTHrP-(1-173) (exon 7) in nonneoplastic tissues. In contrast, a switch in the profile of the different hPTHrP mRNAs was detected in some neoplastic tissues, with an increase in mRNA transcripts encoding PTHrP-(1-173) and a decrease in mRNA transcripts encoding PTHrP-(1-141) observed in several tumors. The differential properties of specific PTHrP 3'-untranslated sequences encoded by unique exons were examined by creating a series of luciferase-PTHrP fusion genes and examining the contribution of PTHrP sequences to relative luciferase activity after transfection of heterologous cell lines. LUC-PTHrP fusion genes containing exon 6 or exon 8 sequences generated consistently higher luciferase activity than fusion genes containing sequences from exon 7. Furthermore, distinct differences in the relative profiles of luciferase activities were observed after transfection of several human, but not rodent, cell lines. These studies suggest that exon splicing at the 3'-end of the hPTHrP gene may be an important determinant of the biological complexity of hPTHrP gene expression.