Prohormone-converting enzymes: regulation and evaluation of function using antisense RNA.
Several putative peptide-processing endoproteases have been identified by homology to the yeast Kex2 endoprotease, including furin, PC2, and PC1. However, the question is still open as to which might be involved in peptide posttranslational processing. To enable detailed comparisons of physiological changes in peptide processing with biochemical and molecular biological studies, we cloned rat pituitary cDNAs for PC1 and PC2. The amino acid sequence homologies among rat, human, and mouse PC1, PC2, and furin are consistent with each being a highly conserved but distinct member of a larger family of mammalian subtilisin-like proteases. PC1 and PC2 mRNAs show a restricted distribution among rat tissues and cultured cell lines, consistent with a role in tissue-specific peptide processing; the occurrence of furin mRNA among these tissues and cell lines is much more widespread, being high in many nonneuroendocrine tissues. In the neurointermediate pituitary, PC1 and PC2 mRNAs are strikingly regulated in response to dopaminergic agents, in parallel with mRNAs for POMC, peptidylglycine alpha-amidating monooxygenase, and carboxypeptidase-H. In AtT-20 cells, PC1 mRNA is coregulated with POMC and peptidylglycine alpha-amidating monooxygenase mRNAs in response to CRH and glucocorticoids. When the endogenous PC1 mRNA level in AtT-20 cells is significantly and specifically decreased by stable expression of antisense RNA to PC1, biosynthetic labeling of newly synthesized POMC-derived peptides shows a substantial blockade of normal POMC processing. These data are consistent with a role for PC1 protein in endoproteolysis, either as a processing endoprotease or as the activator of the actual processing endoprotease(s).
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Neuroscience Department, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
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D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics; D045683:Furin; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics*; D015698:Genomic Library; D006801:Humans; D008297:Male; D051379:Mice; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D010902:Pituitary Gland / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D010911:Pituitary Neoplasms / Q000737:chemistry / Q000235:genetics / Q000473:pathology; D016133:Polymerase Chain Reaction; D011333:Pro-Opiomelanocortin / Q000235:genetics / Q000378:metabolism; D045664:Proprotein Convertase 1; D045706:Proprotein Convertase 2; D043484:Proprotein Convertases; D011498:Protein Precursors / Q000032:analysis / Q000378:metabolism; D016372:RNA, Antisense / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000502:physiology; D051381:Rats; D029701:Saccharomyces cerevisiae Proteins; D012689:Sequence Homology, Nucleic Acid; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism / Q000502:physiology*; D013381:Subtilisins / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000378:metabolism / Q000473:pathology
D009097:Multienzyme Complexes; D011498:Protein Precursors; D016372:RNA, Antisense; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; D011333:Pro-Opiomelanocortin; D004247:DNA; D006899:Mixed Function Oxygenases; C035189:peptidylglycine monooxygenase; D043484:Proprotein Convertases; D012697:Serine Endopeptidases; D013381:Subtilisins; C058751:KEX2 protein, S cerevisiae; D045683:Furin; C473656:Pcsk1 protein, mouse; D045664:Proprotein Convertase 1; D045706:Proprotein Convertase 2