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Measuring Protein Binding to F-actin by Co-sedimentation
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Microvillar and ciliary defects in zebrafish lacking an actin-binding bioactive peptide amidating enzyme

Dhivya Kumar1,2, Rebecca T Thomason3,4, Maya Yankova1,5

  • 1Department of Molecular Biology and Biophysics, University of Connecticut Health Center, Farmington, CT, 06030, USA.

Scientific Reports
|March 16, 2018
PubMed

Insights

Peptidylglycine α-amidating monooxygenase (PAM) is crucial for cell structure. Lack of PAM disrupts microvilli and cilia formation by affecting the actin cytoskeleton, leading to developmental defects.

Area of Science:

  • Cell Biology
  • Developmental Biology
  • Biochemistry

Background:

  • The formation of cellular extensions like microvilli and cilia is vital for cell function but not fully understood.
  • Peptidylglycine α-amidating monooxygenase (PAM) is an enzyme involved in peptide synthesis, found in cilia, and essential for ciliogenesis.
  • Altered PAM levels impact the actin cytoskeleton and secretion in mammalian cells.

Purpose of the Study:

  • To investigate the role of PAM in embryonic development and its impact on microvilli and ciliogenesis.
  • To elucidate the molecular mechanisms by which PAM interacts with the actin cytoskeleton.

Main Methods:

  • Utilized zebrafish (Danio rerio) as a model organism to study the effects of PAM deficiency.
  • Examined microvilli and ciliogenesis in PAM-deficient zebrafish embryos and mouse tracheal epithelial cells.
  • Performed biochemical assays to analyze the interaction between PAM and actin filaments.

Main Results:

  • Zebrafish lacking PAM exhibited a lethal edematous phenotype, loss of microvilli, and impaired ciliogenesis.
  • PAM deficiency in Chlamydomonas led to actin cytoskeleton reorganization and altered actin paralogue expression.
  • The cytosolic C-terminal domain of PAM directly interacts with filamentous actin.

Conclusions:

  • PAM plays a critical role in organizing the actin cytoskeleton during development.
  • This organization by PAM is essential for proper microvillus formation and ciliogenesis.
  • PAM's interaction with actin provides a novel mechanism linking peptide metabolism to cytoskeletal dynamics.

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