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Quantitative Analysis of Cell Edge Dynamics during Cell Spreading
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Arrestins regulate cell spreading and motility via focal adhesion dynamics

Whitney M Cleghorn1, Kevin M Branch2, Seunghyi Kook1

  • 1Department of Pharmacology, Vanderbilt University, Nashville, TN 37232.

Molecular Biology of the Cell
|December 26, 2014
PubMed

Insights

Nonvisual arrestins link microtubules and clathrin, crucial for focal adhesion (FA) disassembly. Lacking arrestins impairs cell motility by increasing FA size and lifespan, highlighting arrestins

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Focal adhesions (FAs) are critical for cell attachment and motility.
  • Timely disassembly of FAs is essential for cell movement.
  • Microtubule-dependent targeting and clathrin recruitment are key to FA disassembly.

Purpose of the Study:

  • To identify molecular links between microtubules and clathrin in FA disassembly.
  • To investigate the role of nonvisual arrestins in regulating focal adhesion dynamics.

Main Methods:

  • Analysis of cells lacking nonvisual arrestins.
  • Cell spreading assays on fibronectin and poly-d-lysine.
  • Nocodazole washout assays to assess microtubule-dependent FA disassembly.
  • Assessment of clathrin dynamics near focal adhesions.
  • Utilizing arrestin mutants deficient in clathrin binding.

Main Results:

  • Cells lacking nonvisual arrestins exhibit excessive spreading, increased adhesion, and reduced motility.
  • Absence of arrestins leads to larger and longer-lived FAs, indicating impaired FA turnover.
  • Arrestin deficiency renders FAs unresponsive to microtubule dynamics, confirming the necessity of arrestins for microtubule-dependent disassembly.
  • Clathrin dynamics near FAs are decreased in arrestin-deficient cells.
  • Arrestin mutants unable to bind clathrin fail to rescue the observed phenotype.

Conclusions:

  • Nonvisual arrestins act as crucial molecular links between microtubules and clathrin.
  • Arrestins are essential regulators of focal adhesion disassembly, controlling FA turnover and cell motility.
  • The clathrin-binding capability of arrestins is vital for their function in focal adhesion regulation.

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