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A more recent version of this article appeared on March 1, 2002
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Submitted on April 17, 2001
Revised on December 5, 2001
Accepted on December 12, 2001
1 Department of Anatomy and Cell Biology, Columbia University
College of Physicians and Surgeons, New York, NY 10032
2 Department of Anatomy and Cell Biology, Columbia University P and S 12-425, 630 W. 168th Street, New York, NY 10032
* Corresponding author. E-mail address: lap5{at}columbia.edu.
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-GFP fusion protein to study motility events associated with inheritance of cortical and nuclear ER in living yeast cells. During M phase prior to nuclear migration, we observe thick, apparently rigid tubular extensions emanating from 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 nuclear ER is inhibited by destabilization of microtubules. Second, astral microtubules, structures that undergo similar patterns of extension, cortical surveillance and retraction, co-localize with nuclear ER extensions. During S and G2 phases of the cell cycle, we observe anchorage of cortical ER at the site of bud emergence and apical bud growth. Thin tubules of ER that extend from the anchored cortical ER display undulating, apparently random movement, and move into the bud as it grows. Finally, we find that cortical ER morphology is sensitive to an F-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.
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