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Vol. 10, Issue 6, 1909-1922, June 1999
School of Biological Sciences, University of Manchester, Manchester
M13 9PT, United Kingdom
The endoplasmic reticulum (ER) in animal cells uses microtubule
motor proteins to adopt and maintain its extended, reticular organization. Although the orientation of microtubules in many somatic
cell types predicts that the ER should move toward microtubule plus
ends, motor-dependent ER motility reconstituted in extracts of
Xenopus laevis eggs is exclusively a minus end-directed,
cytoplasmic dynein-driven process. We have used Xenopus
egg, embryo, and somatic Xenopus tissue culture cell
(XTC) extracts to study ER motility during embryonic development in
Xenopus by video-enhanced differential interference
contrast microscopy. Our results demonstrate that cytoplasmic dynein is
the sole motor for microtubule-based ER motility throughout the early
stages of development (up to at least the fifth embryonic interphase).
When egg-derived ER membranes were incubated in somatic XTC cytosol,
however, ER tubules moved in both directions along microtubules. Data
from directionality assays suggest that plus end-directed ER tubule
extensions contribute ~19% of the total microtubule-based ER
motility under these conditions. In XTC extracts, the rate of ER tubule
extensions toward microtubule plus ends is lower (~0.4 µm/s) than
minus end-directed motility (~1.3 µm/s), and plus end-directed
motility is eliminated by a function-blocking anti-conventional kinesin
heavy chain antibody (SUK4). In addition, we provide evidence that the
initiation of plus end-directed ER motility in somatic cytosol is
likely to occur via activation of membrane-associated kinesin.
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