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TTL-Expression Modulates Epithelial Morphogenesis.

Manuel Müller1,2, Karina Ringer1,2, Florian Hub3

  • 1Department of Cell Biology and Cell Pathology, Philipps-Universität Marburg, Marburg, Germany.

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|February 22, 2021
PubMed
Summary
This summary is machine-generated.

Tubulin tyrosination, regulated by tubulin tyrosine ligase (TTL), impacts epithelial cell shape and adhesion. Loss of TTL alters microtubule composition, affecting cell morphology and focal adhesion dynamics crucial for tissue formation.

Keywords:
epithelia cellsfocal adhesionintestinal organoidmicrotubule tyrosination/detyrosinationtubulin tyrosine ligase

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Area of Science:

  • Cell Biology
  • Cytoskeletal Dynamics
  • Epithelial Biology

Background:

  • Epithelial monolayer formation relies on microtubule cytoskeleton architecture and composition.
  • Microtubules regulate intracellular trafficking and positioning of cellular proteins.
  • Tubulin tyrosination is a post-translational modification influencing microtubule function.

Purpose of the Study:

  • To investigate the role of tubulin tyrosination, specifically the enzyme tubulin tyrosine ligase (TTL), in epithelial cell shape, motility, and adhesion.
  • To elucidate how altering TTL levels affects microtubule composition and downstream cellular processes.
  • To understand the impact of detyrosinated versus tyrosinated tubulin equilibrium on epithelial tissue formation.

Main Methods:

  • Depletion and overexpression of tubulin tyrosine ligase (TTL) in 2D epithelial monolayers and 3D intestinal organoids.
  • Qualitative and quantitative analysis of cell morphology and shape acquisition.
  • Assessment of cell adhesion patterns and migration dynamics.
  • Immunoprecipitation to analyze the association of detyrosinated tubulin with focal adhesion components.

Main Results:

  • Absence of TTL led to high levels of detyrosinated tubulin, initial flat cell morphology, and delayed acquisition of columnar shape.
  • TTL-knockout cells exhibited enhanced adhesion and accelerated migration.
  • TTL overexpression reversed these effects, indicating TTL's critical role.
  • Detyrosinated tubulin co-precipitated with focal adhesion scaffold components, increasing focal adhesion plaque quantity and persistence.

Conclusions:

  • The equilibrium between detyrosinated and tyrosinated tubulin modulates epithelial tissue formation.
  • Tubulin tyrosination status significantly impacts epithelial cell morphology and adhesion dynamics.
  • TTL activity is a key regulator of cytoskeletal organization and cell-matrix interactions in epithelial tissues.