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Vol. 8, Issue 10, 1971-1988, October 1997
and
Departments of
*Morphology and
The discovery that the dilute gene encodes a class V
myosin led to the hypothesis that this molecular motor is involved in melanosome transport and/or dendrite outgrowth in mammalian
melanocytes. The present studies were undertaken to gain insight into
the subcellular distribution of myosin-V in the melanoma cell line
B16-F10, which is wild-type for the dilute gene.
Immunofluorescence studies showed some degree of superimposed labeling
of myosin-V with melanosomes that predominated at the cell periphery. A
subcellular fraction highly enriched in melanosomes was also enriched
in myosin-V based on Western blot analysis. Immunoelectron microscopy
showed myosin-V labeling associated with melanosomes and other
organelles. The stimulation of B16 cells with the
Biochemistry,
Faculdade de Medicina de Ribeirão Preto-Universidade de São
Paulo, Ribeirão Preto, São Paulo, 14049-900, and
Faculty of Farmaceutic Sciences-Universidade Federal de
Goiás, 74021-070 Goiânia, Goiás, Brazil
-melanocyte-stimulating hormone led to a significant increase in
myosin-V expression. This is the first evidence that a cAMP signaling
pathway might regulate the dilute gene expression.
Immunofluorescence also showed an intense labeling of myosin-V
independent of melanosomes that was observed within the dendrites and
at the perinuclear region. Although the results presented herein are
consistent with the hypothesis that myosin-V might act as a motor for
melanosome translocation, they also suggest a broader cytoplasmic
function for myosin-V, acting on other types of organelles or in
cytoskeletal dynamics.
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