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Aip1p Dynamics Are Altered by the R256H Mutation in Actin
Published on: July 30, 2014
Arp2/3 complex and actin dynamics are required for actin-based mitochondrial motility in yeast
I R Boldogh1, H C Yang, W D Nowakowski
1Department of Anatomy and Cell Biology, Columbia University, New York, NY 10032, USA.
Abstract:
The Arp2/3 complex is implicated in actin polymerization-driven movement of Listeria monocytogenes. Here, we find that Arp2p and Arc15p, two subunits of this complex, show tight, actin-independent association with isolated yeast mitochondria. Arp2p colocalizes with mitochondria. Consistent with this result, we detect Arp2p-dependent formation of actin clouds around mitochondria in intact yeast. Cells bearing mutations in ARP2 or ARC15 genes show decreased velocities of mitochondrial movement, loss of all directed movement and defects in mitochondrial morphology. Finally, we observe a decrease in the velocity and extent of mitochondrial movement in yeast in which actin dynamics are reduced but actin cytoskeletal structure is intact. These results support the idea that the movement of mitochondria in yeast is actin polymerization driven and that this movement requires Arp2/3 complex.
Insights
The Arp2/3 complex drives actin polymerization for Listeria movement. In yeast, this complex is crucial for actin-driven mitochondrial movement, affecting velocity and morphology.
Area of Science:
- Cell Biology
- Molecular Biology
- Cytoskeletal Dynamics
Background:
- The Arp2/3 complex is known to regulate actin polymerization, crucial for processes like cell motility and intracellular pathogen movement.
- Mitochondrial dynamics and movement are essential for cellular energy homeostasis and function, but the precise mechanisms are still being elucidated.
Purpose of the Study:
- To investigate the role of the Arp2/3 complex in the movement of mitochondria within yeast cells.
- To determine if actin polymerization is the driving force behind mitochondrial transport in yeast.
Main Methods:
- Co-immunoprecipitation to assess the association between Arp2/3 subunits and mitochondria.
- Fluorescence microscopy to visualize the localization of Arp2p and actin structures around mitochondria.
- Analysis of mitochondrial movement velocity and morphology in wild-type and mutant yeast strains (ARP2, ARC15) and under conditions of reduced actin dynamics.
Main Results:
- The Arp2p and Arc15p subunits of the Arp2/3 complex associate with yeast mitochondria independently of actin.
- Arp2p colocalizes with mitochondria and forms actin clouds around them in intact yeast cells.
- Mutations in ARP2 or ARC15, or impaired actin dynamics, lead to reduced mitochondrial movement velocity, loss of directed movement, and morphological defects.
Conclusions:
- Mitochondrial movement in yeast is driven by actin polymerization.
- The Arp2/3 complex is essential for actin-driven mitochondrial motility and proper mitochondrial morphology in yeast.
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