Targeting of pro-opiomelanocortin to the regulated secretory pathway may involve cooperation between different protein domains.
The structure of pro-opiomelanocortin (POMC) can be divided into three main domains: an NH2-terminal domain formed by the NH2-terminal glycopeptide and the joining peptide, a central domain corresponding to the adrenocorticotropin sequences and a COOH-terminal domain containing the beta-lipotropin sequences. Expression of POMC in neuroendocrine cell lines such as the mouse neuroblastoma Neuro2A cells results in its targeting to the regulated secretory pathway of these cells. Intracellular targeting of proteins along non default pathways are widely believed to involve the recognition of specific structural features by a sorting machinery. To understand the nature of the signal involved in targeting prohormone to the regulated secretory pathway, we have constructed mutants of POMC in which sequences from the NH2-terminal, the central and the COOH-terminal domains were deleted and examined the sorting of these mutant POMC molecules in Neuro2A cells by immunofluorescence and immunoelectron microscopy. Our results indicate that POMC NH2-terminal glycopeptide or beta-LPH domain do not contain sufficient information for targeting to the regulated pathway since these peptides are not sorted to secretory vesicles when expressed in Neuro2A cells: Similarly, the ACTH domain does not contain essential targeting information since POMC mutants lacking these sequences were sorted to secretory vesicles. Mutant POMCs containing the sequences of more than one of the main protein domains were, however, correctly targeted to the regulated secretory pathway. Our results indicate that POMC is not targeted to the regulated secretory pathway through recognition of a unique continuous 'molecular address'.(ABSTRACT TRUNCATED AT 250 WORDS)
Chevrier
D
D
Département de Biochimie, Faculté de Médecine, Université de Montréal, Canada.
Fournier
H
H
Nault
C
C
Zollinger
M
M
Crine
P
P
Boileau
G
G
D000324:Adrenocorticotropic Hormone / Q000032:analysis / Q000378:metabolism; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002461:Cell Line, Transformed; D004247:DNA / Q000032:analysis / Q000235:genetics; D005455:Fluorescent Antibody Technique; D017353:Gene Deletion; D015151:Immunoblotting; D051379:Mice; D016253:Microscopy, Immunoelectron; D008969:Molecular Sequence Data; D009154:Mutation; D009447:Neuroblastoma / Q000737:chemistry / Q000473:pathology / Q000648:ultrastructure; D011333:Pro-Opiomelanocortin / Q000032:analysis* / Q000235:genetics / Q000502:physiology*; D011863:Radioimmunoassay; D014407:Tumor Cells, Cultured
D000596:Amino Acids; D011333:Pro-Opiomelanocortin; D000324:Adrenocorticotropic Hormone; D004247:DNA