Melanogenesis in transfected fibroblasts induces lysosomal activation.
Normal melanosome biogenesis requires the association of structural proteins with tyrosinase. 3T3 Swiss fibroblasts transfected with mouse tyrosinase cDNA (line 13.4, clone c) are a unique system in which melanogenesis takes place in the absence of melanosomal structural proteins. Our study confirmed that transfected fibroblasts displayed tyrosinase activity and some of them produced pigment granules. In the absence of melanosomal structural proteins the granules failed both to show a typical ultrastructure and to undergo the usual melanosome ontogenesis. The differentiating agent--dimethyl sulfoxide--increased phaeomelanin production. Pigment was localized in membrane-bound vesicles which were identified as lysosomes by means of immunogold electron microscopy. Cell line 13.4 had higher levels of lysosomal enzymes (beta-hexosaminidase, alpha-mannosidase) than both parental 3T3 cells and clone pKG4 (fibroblasts transfected with the G418 resistance plasmid). Melanosomal proteins act as scavengers of toxic products of melanogenesis, and our results suggest that in their absence cells may employ an alternative mechanism to sequester injurious products.
Borovanský
J
J
Department of Dermatology, University Hospital, Leiden, The Netherlands.
Mommaas
A M
AM
Smit
N P
NP
Eygendaal
D
D
Winder
A J
AJ
Vermeer
B J
BJ
Pavel
S
S
D016475:3T3 Cells; D000818:Animals; D002999:Clone Cells; D018076:DNA, Complementary; D004798:Enzymes / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism* / Q000648:ultrastructure; D008247:Lysosomes / Q000378:metabolism / Q000502:physiology* / Q000648:ultrastructure; D008543:Melanins / Q000096:biosynthesis*; D051379:Mice; D008854:Microscopy, Electron; D014442:Monophenol Monooxygenase / Q000235:genetics / Q000378:metabolism; D010860:Pigments, Biological / Q000378:metabolism; D014162:Transfection
D018076:DNA, Complementary; D004798:Enzymes; D008543:Melanins; D010860:Pigments, Biological; D014442:Monophenol Monooxygenase