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Cloning and Primary Sequence of a Mouse Candidate Prohormone Convertase PC1 Homologous to PC2, Furin, and Kex2: Distinct Chromosomal Localization and Messenger RNA Distribution in Brain and Pituitary Compared to PC2

J. A. DeSève Laboratories of Biochemical, Clinical Research Institute of Montreal Montreal, Quebec, H2W 1R7 Canada (Affiliated with I'Universiteé de Montréal)
Molecular Neuroendocrinology, Clinical Research Institute of Montreal Montreal, Quebec, H2W 1R7 Canada (Affiliated with I'Universiteé de Montréal)
Hôpital d'Enfants Groupe Hospitalier de la Timone 13385 Marseille Cedex 5, France

Address requests for reprints to: Dr. N. G. Seidah, J. A. DeSeve Laboratories of Biochemical Neuroendrocrinology, Clinical Research Institute of Montreal, 110 Pine Avenue West, Montreal, Quebec, H2W 1R7 Canada.

Abstract

Using a 796-basepair cDNA fragment obtained from a mouse pituitary library we have screened two mouse insulinoma libraries and isolated a full-length cDNA clone (2516 basepairs; 753 amino acids), designated mPC1. The cDNA sequence of mPC1 codes for a protein containing 753 amino acids and three potential N-glycosylation sites. This cDNA encodes a putative novel subtilisin-like proteinase, exhibiting within its presumed catalytic domain 64%, 55%, and 47% amino acid sequence identity to the recently characterized candidate prohormone convertases human Furin, mouse PC2, and yeast Kex2 gene products, respectively. An identical sequence to mPC1 was derived from a cDNA library of mouse corticotroph AtT-20 tumor cells. An ArgGlyAsp tripeptide identical to the recognition sequence of integrins was observed in the structures of the mammalian PC1, PC2, and Furin. In situ hybridization results demonstrated a distinct localization of the mPC1 and mPC2 transcripts in pituitary and brain. Thus, whereas both mPC1 and mPC2 are found in the intermediate lobe of the pituitary, only mPC1 is easily detected in the anterior lobe. In extrahypothalamic regions of the brain, including cortex, hippocampus, thalamus, and spinal cord, mPC2 transcripts predominate over mPC1. Both mRNAs are found in only a fraction of hypothalamic neurons, with greater abundance of mPC1 over mPC2 in the supraoptic nucleus. The genes coding for mPC1 and mPC2 map to the murine chromosomes 13 (band 13c) and 2 (2F3–2H2 region), respectively.

FOOTNOTES

This work was supported by a research grant from the Medical Research Council of Canada and J. A. de Seve Succession.

^* On leave from Endocrine Unit, First Department of Medicine, Szeged University Medical School, Szeged, Hungary.

Received for publication August 6, 1990. Revision received October 26, 1990. Accepted for publication October 26, 1990.

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