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J. Cell Biol.,
Volume 143, Number 6, December 14, 1998 1547-1558


* Department of Cell and Structural Biology, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801; Melanophores move pigment organelles
(melanosomes) from the cell center to the periphery
and vice-versa. These bidirectional movements require
cytoplasmic microtubules and microfilaments and depend on the function of microtubule motors and a myosin. Earlier we found that melanosomes purified from
Xenopus melanophores contain the plus end microtubule motor kinesin II, indicating that it may be involved
in dispersion (Rogers, S.L., I.S. Tint, P.C. Fanapour,
and V.I. Gelfand. 1997. Proc. Natl. Acad. Sci. USA. 94:
3720-3725). Here, we generated a dominant-negative
construct encoding green fluorescent protein fused to
the stalk-tail region of Xenopus kinesin-like protein 3 (Xklp3), the 95-kD motor subunit of Xenopus kinesin
II, and introduced it into melanophores. Overexpression of the fusion protein inhibited pigment dispersion
but had no effect on aggregation. To control for the
specificity of this effect, we studied the kinesin-dependent movement of lysosomes. Neither dispersion of lysosomes in acidic conditions nor their clustering under
alkaline conditions was affected by the mutant Xklp3.
Furthermore, microinjection of melanophores with
SUK4, a function-blocking kinesin antibody, inhibited
dispersion of lysosomes but had no effect on melanosome transport. We conclude that melanosome dispersion is powered by kinesin II and not by conventional
kinesin. This paper demonstrates that kinesin II moves
membrane-bound organelles.
European
Molecular Biology Laboratory (EMBL), Heidelberg, Germany, D-69117; and § Department of Anatomy, School of Medicine,
Indiana University, Indianapolis, Indiana 46202
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