Differential localization and dynamics of class I myosins in the enterocyte microvillus

Mol Biol Cell. 2010 Mar 15;21(6):970-8. doi: 10.1091/mbc.e09-07-0638. Epub 2010 Jan 20.

Abstract

Epithelial cells lining the intestinal tract build an apical array of microvilli known as the brush border. Each microvillus is a cylindrical membrane protrusion that is linked to a supporting actin bundle by myosin-1a (Myo1a). Mice lacking Myo1a demonstrate no overt physiological symptoms, suggesting that other myosins may compensate for the loss of Myo1a in these animals. To investigate changes in the microvillar myosin population that may limit the Myo1a KO phenotype, we performed proteomic analysis on WT and Myo1a KO brush borders. These studies revealed that WT brush borders also contain the short-tailed class I myosin, myosin-1d (Myo1d). Myo1d localizes to the terminal web and striking puncta at the tips of microvilli. In the absence of Myo1a, Myo1d peptide counts increase twofold; this motor also redistributes along the length of microvilli, into compartments normally occupied by Myo1a. FRAP studies demonstrate that Myo1a is less dynamic than Myo1d, providing a mechanistic explanation for the observed differential localization. These data suggest that Myo1d may be the primary compensating class I myosin in the Myo1a KO model; they also suggest that dynamics govern the localization and function of different yet closely related myosins that target common actin structures.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • Enterocytes / cytology*
  • Fluorescence Recovery After Photobleaching
  • Mice
  • Mice, Knockout
  • Microvilli / metabolism*
  • Microvilli / ultrastructure
  • Myosin Heavy Chains / genetics
  • Myosin Heavy Chains / metabolism*
  • Myosins / genetics
  • Myosins / metabolism*
  • Protein Isoforms / genetics
  • Protein Isoforms / metabolism
  • Proteomics / methods
  • Rats
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Myo1a protein, mouse
  • Protein Isoforms
  • Recombinant Fusion Proteins
  • Myo1d protein, mouse
  • Myosin Heavy Chains
  • Myosins