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Related Experiment Videos

Thy-1 expresses two signals for apical localization in epithelial cells.

S K Powell1, M P Lisanti, E J Rodriguez-Boulan

  • 1Department of Cell Biology and Anatomy, Cornell University Medical College, New York, New York 10021.

The American Journal of Physiology
|April 1, 1991
PubMed
Summary

Glycosyl phosphatidylinositol (GPI) anchoring directs proteins to the apical surface in epithelial cells. This study shows Thy-1 protein also contains apical targeting information within its sequence, independent of GPI anchoring.

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Area of Science:

  • Cell biology
  • Molecular biology
  • Protein targeting

Background:

  • Glycosyl phosphatidylinositol (GPI) anchoring is known to mediate apical targeting of plasma membrane proteins in epithelial cells.
  • The precise mechanisms and protein-specific signals for apical localization are still under investigation.

Purpose of the Study:

  • To investigate the role of GPI anchoring and protein sequence in the apical targeting of Thy-1.
  • To determine if Thy-1 possesses intrinsic apical targeting signals.

Main Methods:

  • Transfection of Madin-Darby canine kidney cells with GPI-anchored and secretory forms of Thy-1.
  • Expression analysis of wild-type and truncated Thy-1 constructs.
  • Localization studies using immunofluorescence.

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Main Results:

  • Native GPI-anchored Thy-1 was localized to the apical surface of transfected cells.
  • A truncated Thy-1 mutant, unable to attach a GPI anchor, was secreted to the apical surface.
  • These findings suggest dual apical targeting mechanisms for Thy-1.

Conclusions:

  • Thy-1 contains specific protein sequence elements that direct it to the apical surface.
  • Both the GPI anchor and the protein sequence contribute to the apical localization of Thy-1.
  • This dual targeting mechanism highlights the complexity of protein sorting in epithelial cells.