Normal tyrosine transport and abnormal tyrosinase routing in pink-eyed dilution melanocytes.
The pink-eyed dilution phenotype in mice arises from mutations in the p gene; in humans, analogous mutations in the P gene result in oculocutaneous albinism type 2. Although the molecular mechanisms which underlie this phenotype remain obscure, it has been postulated that mutations in p result in defective tyrosine transport into murine melanosomes, resulting in hypopigmentation and diminished coat color. However, we previously reported no difference in melanosomal tyrosine transport in unpigmented, melanoblast-like pink-eyed dilution (pcp/pcp), and in pigmented (melan-a) murine melanocytes. In this study, we utilized melan-p1 cells, more differentiated pink-eyed dilution (pcp/p25H) melanocytes which can be induced to produce melanin, to characterize the melanogenic lesion(s) more definitively. Uptake of [3H]tyrosine into melan-a melanosomes did not differ significantly from uptake into melanosomes derived from melan-p1 melanocytes, further arguing against its critical role as a tyrosine transporter. Pink-eyed dilution melanocytes incubated in high tyrosine concentrations became extremely pigmented as they became confluent and secreted large amounts of black material into the medium. Total cellular tyrosinase activity in melan-p1 melanocytes was significantly higher than that in melan-a melanocytes (which are wild-type at the p locus), but the localization of tyrosinase to melanosomes was impaired in melan-p1 melanocytes compared to melan-a melanocytes. These results indicate that mechanisms other than deficient tyrosine transport are involved in the pink-eyed dilution phenotype and that this protein may serve a chaperone-like or stabilizing function in melanocytes.
Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, 20892, USA.
S B
SB
Potterf
M
M
Furumura
E V
EV
Sviderskaya
C
C
Santis
D C
DC
Bennett
V J
VJ
Hearing
D016115:Albinism, Oculocutaneous / Q000235:genetics; D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins; D002460:Cell Line; D008543:Melanins / Q000096:biosynthesis; D008544:Melanocytes / Q000201:enzymology / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008810:Mice, Inbred C57BL; D014442:Monophenol Monooxygenase / Q000378:metabolism*; D009154:Mutation; D010641:Phenotype; D010858:Pigmentation / Q000235:genetics; D013347:Subcellular Fractions / Q000201:enzymology; D014443:Tyrosine / Q000378:metabolism* / Q000502:physiology
D002352:Carrier Proteins; D008543:Melanins; D008565:Membrane Proteins; C076239:P protein, mouse; D014443:Tyrosine; D014442:Monophenol Monooxygenase