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Vol. 13, Issue 3, 854-865, March 2002
Department of Anatomy and Cell Biology, Columbia
University, College of Physicians and Surgeons, New York, New York
10032
The endoplasmic reticulum (ER) in Saccharomyces
cerevisiae consists of a reticulum underlying the plasma
membrane (cortical ER) and ER associated with the nuclear envelope
(nuclear ER). We used a Sec63p-green fluorescent protein fusion protein
to study motility events associated with inheritance of cortical ER and nuclear ER in living yeast cells. During M phase before nuclear migration, we observed thick, apparently rigid tubular extensions emanating from the nuclear ER that elongate, undergo sweeping motions
along the cell cortex, and shorten. Two findings support a role for
microtubules in this process. First, extension of tubular structures
from the nuclear ER is inhibited by destabilization of microtubules.
Second, astral microtubules, structures that undergo similar patterns
of extension, cortical surveillance and retraction, colocalize with
nuclear ER extensions. During S and G2 phases of the cell
cycle, we observed anchorage of the cortical ER at the site of bud
emergence and apical bud growth. Thin tubules of the ER that extend
from the anchored cortical ER display undulating, apparently random
movement and move into the bud as it grows. Finally, we found that
cortical ER morphology is sensitive to a filamentous
actin-destabilizing drug, latrunculin-A, and to mutations in the
actin-encoding ACT1 gene. Our observations support 1)
different mechanisms and cytoskeletal mediators for the inheritance of
nuclear and cortical ER elements and 2) a mechanism for cortical ER
inheritance that is cytoskeleton dependent but relies on anchorage, not
directed movement.
Online version of this article contains video material. Online
version is available at www.molbiolcell.org.
*
Corresponding author. E-mail address:
lap5{at}columbia.edu.
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