Endoplasmic reticulum dynamics, inheritance, and cytoskeletal interactions in budding yeast

Mol Biol Cell. 2002 Mar;13(3):854-65. doi: 10.1091/mbc.01-04-0184.

Abstract

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 G(2) 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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Actin Cytoskeleton / metabolism
  • Bridged Bicyclo Compounds, Heterocyclic / metabolism
  • Cell Cycle / physiology
  • Cytoskeleton / metabolism*
  • Dimethyl Sulfoxide / metabolism
  • Endoplasmic Reticulum / metabolism*
  • Fungal Proteins / genetics
  • Fungal Proteins / metabolism*
  • Genes, Reporter
  • Heat-Shock Proteins*
  • Intracellular Membranes / metabolism
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Membrane Transport Proteins*
  • Microtubules / metabolism
  • Models, Biological
  • Nuclear Envelope / metabolism*
  • Recombinant Fusion Proteins / metabolism
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae / ultrastructure
  • Saccharomyces cerevisiae Proteins*
  • Solvents / metabolism
  • Thiazoles / metabolism
  • Thiazolidines

Substances

  • Bridged Bicyclo Compounds, Heterocyclic
  • Fungal Proteins
  • Heat-Shock Proteins
  • Membrane Proteins
  • Membrane Transport Proteins
  • Recombinant Fusion Proteins
  • SEC63 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Solvents
  • Thiazoles
  • Thiazolidines
  • latrunculin A
  • Dimethyl Sulfoxide