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Calcium Research Laboratory, Departments of Physiology and Medicine, McGill University and Royal Victoria Hospital Montreal, Canada H3A 1A1
Laboratory of Molecular Genetics, National Research Council, Biotechnology Research Institute Montreal, Canada H4P 2R2
Address requests for reprints to: Dr. David Goltzman, Calcium Research Laboratory, Royal Victoria Hospital, Room H4.67, 687 Pine Avenue West, Montreal, Canada H3A 1A1.
Abstract
A rat Leydig cell tumor cDNA library was screened with a 32P-labeled genomic restriction fragment encoding human PTH-like peptide (hPLP), and three cDNA clones were isolated. The largest cDNA insert contained 1146 nucleotides. The cloned cDNA encodes a 177-amino acid protein consisting of a predicted 36-amino acid leader sequence and a 141-amino acid mature peptide in which 9 of 13 aminoterminal residues are identical to rat PTH (rPTH). Comparison of rPLP with hPLP reveals marked conservation of both the nucleotide and amino acid sequences through the prepro, amino-terminal, and midregion portions of the molecules. There is also striking conservation of the 3' noncoding regions of rPLP and hPLP mRNAs, both of which contain AU-rich repeated sequences that may affect mRNA stability. A single species of mRNA of approximately 1.4-kilobases was identified in the rat Leydig cell tumor and in normal rat stomach. Southern blot analysis is consistent with the presence of a single copy of the rPLP gene per haploid genome, and there is no major rearrangement or amplification of the rPLP gene in DNA isolated from the tumor per se. The results demonstrate the presence of a single gene transcript in a rat model of malignancy associated with hypercalcemia which encodes a peptide homologous to hPLP, document the marked interspecies sequence conservation that exists in major functional domains of the mRNAs and peptides, and show the expression of mRNA encoding rPLP in normal stomach as well as in neoplastic rat tissue.
FOOTNOTES
This work was supported by a grant from the NCI of Canada and Grants MT-5775 and MA-9315 from the Medical Research Council (MRC) of Canada.
* Recipient of a fellowship from the MRC of Canada.
Recipient of a Scholarship from the MRC of Canada.
Received for publication November 18, 1988. Revision received December 13, 1988. Accepted for publication December 14, 1988.
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