Characterization of the cDNA encoding the prohormone convertase PC2 and localization of the mRNA in the brain of the frog Rana ridibunda.
A number of precursors for neuropeptides have recently been cloned in amphibians, but little is known concerning the endoproteases responsible for the processing of these precursors. Here we report on the molecular cloning of the cDNA encoding the proprotein convertase PC2 and the distribution of the corresponding mRNA in the European green frog Rana ridibunda. The full cDNA structure (2125 bp) was obtained from the analysis of the PCR products combined with the sequence from a clone isolated from a frog pituitary cDNA library. The deduced amino acid sequence revealed that frog PC2 comprises 636 amino acid residues including a 22-residue signal peptide. RT-PCR analysis showed that PC2 is expressed not only in the brain and pituitary but also in various peripheral organs including the pancreas, stomach, intestine, liver, kidney and testis. In situ hybridization histochemistry revealed that, in the central nervous system, PC2 mRNA is widely distributed, the highest concentrations being found in the pallium, the anterior preoptic area, the hypothalamus and the medial amygdala. High levels of PC2 mRNA were also detected in the intermediate lobe of the pituitary. The overall distribution of PC2 mRNA in the frog brain is consistent with its involvement in the processing of a number of neuropeptide and hormone precursors.
Vieau
D
D
European Institute for Peptide Research (IFRMP no. 23), Laboratory of Cellular and Molecular Neuroendocrinology, INSERM U413, UA CNRS, University of Rouen, 76821 Mont-Saint-Aignan, France.
Gangnon
F
F
Jégou
S
S
Danger
J M
JM
Vaudry
H
H
D000818:Animals; D016375:Antisense Elements (Genetics); D001483:Base Sequence; D001921:Brain / Q000201:enzymology*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D017403:In Situ Hybridization; D008297:Male; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000378:metabolism; D010902:Pituitary Gland / Q000737:chemistry; D045706:Proprotein Convertase 2; D012333:RNA, Messenger / Q000032:analysis; D011895:Rana ridibunda; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013381:Subtilisins / Q000235:genetics*
D016375:Antisense Elements (Genetics); D018076:DNA, Complementary; D009479:Neuropeptides; D012333:RNA, Messenger; D013381:Subtilisins; D045706:Proprotein Convertase 2