Characterization of human palladin, a microfilament-associated protein

Mol Biol Cell. 2001 Oct;12(10):3060-73. doi: 10.1091/mbc.12.10.3060.

Abstract

Actin-containing microfilaments control cell shape, adhesion, and contraction. In striated muscle, alpha-actinin and other Z-disk proteins coordinate the organization and functions of actin filaments. In smooth muscle and nonmuscle cells, periodic structures termed dense bodies and dense regions, respectively, are thought to serve functions analogous to Z-discs. We describe here identification and characterization of human palladin, a protein expressed mainly in smooth muscle and nonmuscle and distributed along microfilaments in a periodic manner consistent with dense regions/bodies. Palladin contains three Ig-domains most homologous to the sarcomeric Z-disk protein myotilin. The N terminus includes an FPPPP motif recognized by the Ena-Vasp homology domain 1 domain in Ena/vasodilatator-stimulated phosphoprotein (VASP)/Wiscott-Aldrich syndrome protein (WASP) protein family. Cytoskeletal proteins with FPPPP motif target Ena/VASP/WASP proteins to sites of actin modulation. We identified palladin in a yeast two-hybrid search as an ezrin-associated protein. An interaction between palladin and ezrin was further verified by affinity precipitation and blot overlay assays. The interaction was mediated by the alpha-helical domain of ezrin and by Ig-domains 2-3 of palladin. Ezrin is typically a component of the cortical cytoskeleton, but in smooth muscle cells it is localized along microfilaments. These cells express palladin abundantly and thus palladin may be involved in the microfilament localization of ezrin. Palladin expression was up-regulated in differentiating dendritic cells (DCs), coinciding with major cytoskeletal and morphological alterations. In immature DCs, palladin localized in actin-containing podosomes and in mature DCs along actin filaments. The regulated expression and localization suggest a role for palladin in the assembly of DC cytoskeleton.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actin Cytoskeleton / chemistry
  • Actin Cytoskeleton / metabolism*
  • Amino Acid Sequence
  • Cell Differentiation / physiology
  • Cells, Cultured
  • Cloning, Molecular / methods
  • Cytoskeletal Proteins / chemistry*
  • Cytoskeletal Proteins / metabolism*
  • Cytoskeletal Proteins / ultrastructure
  • DNA, Complementary / chemistry
  • DNA, Complementary / metabolism
  • Dendritic Cells / cytology
  • Glioma
  • HeLa Cells
  • Humans
  • Immunohistochemistry / methods
  • Microscopy, Fluorescence / methods
  • Molecular Sequence Data
  • Phosphoproteins / chemistry*
  • Phosphoproteins / metabolism*
  • Phosphoproteins / ultrastructure
  • RNA, Messenger / chemistry
  • RNA, Messenger / metabolism*
  • Sequence Analysis / methods
  • Subcellular Fractions / chemistry
  • Subcellular Fractions / metabolism
  • Tumor Cells, Cultured

Substances

  • Cytoskeletal Proteins
  • DNA, Complementary
  • PALLD protein, human
  • Phosphoproteins
  • RNA, Messenger
  • ezrin