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Related Experiment Videos

Actin and septin ultrastructures at the budding yeast cell cortex.

Avital A Rodal1, Lukasz Kozubowski, Bruce L Goode

  • 1Department of Biology, Rosenstiel Basic Medical Science Research Center, Brandeis University, Waltham, MA 02454, USA. arodal@mit.edu

Molecular Biology of the Cell
|November 5, 2004
PubMed
Summary

Budding yeast actin and septin organization at the cell cortex was visualized using rapid-freeze and deep-etch techniques. This study reveals new details about actin patch assembly during endocytosis and septin filament higher-order structures.

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Area of Science:

  • Cell Biology
  • Cytoskeleton Dynamics
  • Microscopy Techniques

Background:

  • Budding yeast is a model organism for studying actin in endocytosis and septins in cell division.
  • Lack of ultrastructural data has limited mechanistic understanding of actin and septin organization at the cell cortex.

Purpose of the Study:

  • To provide high-resolution ultrastructural information on the organization of actin and septins at the yeast cell cortex.
  • To elucidate the roles of actin and septins in endocytosis and cell division.

Main Methods:

  • Development and application of rapid-freeze and deep-etch techniques for imaging.
  • High-resolution imaging of the yeast cell cortex in spheroplasted cells.
  • Analysis of actin and septin structures in wild-type and mutant yeast cells.

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Main Results:

  • Cortical actin cytoskeleton forms conical or mound-like structures with short, cross-linked filaments.
  • Arp2/3 complex localizes at the apex of actin structures, suggesting initiation of actin patch assembly.
  • Septin filaments assemble into rings and ordered gauzes at the cell cortex.
  • Mutants affecting endocytosis display defects in cortical actin patch assembly.

Conclusions:

  • A model for actin function during endocytosis is proposed based on observed structures and mutant phenotypes.
  • The study provides the first high-resolution views of septin organization within cells.
  • Ultrastructural insights advance understanding of cytoskeletal roles in yeast cell processes.