Transforming growth factor-β receptors interact with AP2 by direct binding to β2 subunit

Diying Yao, Marcelo Ehrlich, Yoav I. Henis, Edward B Leof

Research output: Contribution to journalArticle

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Abstract

Transforming growth factor-β (TGF-β) superfamily members regulate a wide range of biological processes by binding to two transmembrane serine/threonine kinase receptors, type I and type II. We have previously shown that the internalization of these receptors is inhibited by K+ depletion, cytosol acidification, or hypertonic medium, suggesting the involvement of clathrin-coated pits. However, the involvement of the clathrin-associated adaptor complex AP2 and the identity of the AP2 subunit that binds the receptors were not known. Herein, we have studied these issues by combining studies on intact cells with in vitro assays. Using fluorescence photobleaching recovery to measure the lateral mobility of the receptors on live cells (untreated or treated to alter their coated pit structure), we demonstrated that their mobility is restricted by interactions with coated pits. These interactions were transient and mediated through the receptors' cytoplasmic tails. To measure direct binding of the receptors to specific AP2 subunits, we used yeast two-hybrid screens and in vitro biochemical assays. In contrast to most other plasma membrane receptors that bind to AP2 via the μ2 subunit, AP2/TGF-β receptor binding was mediated by a direct interaction between the β2-adaptin N-terminal trunk domain and the cytoplasmic tails of the receptors; no binding was observed to the μ2, α, or σ2 subunits of AP2 or to μ1 of AP1. The data uniquely demonstrate both in vivo and in vitro the ability of β2-adaptin to directly couple TGF-β receptors to AP2 and to clathrin-coated pits, providing the first in vivo evidence for interactions of a transmembrane receptor with β2-adaptin.

Original languageEnglish (US)
Pages (from-to)4001-4012
Number of pages12
JournalMolecular Biology of the Cell
Volume13
Issue number11
DOIs
StatePublished - Nov 1 2002

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Growth Factor Receptors
Transforming Growth Factors
Clathrin
Cytoplasmic and Nuclear Receptors
Vesicular Transport Adaptor Proteins
Fluorescence Recovery After Photobleaching
Biological Phenomena
Protein-Serine-Threonine Kinases
Cytosol
Yeasts
Cell Membrane
In Vitro Techniques

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Transforming growth factor-β receptors interact with AP2 by direct binding to β2 subunit. / Yao, Diying; Ehrlich, Marcelo; Henis, Yoav I.; Leof, Edward B.

In: Molecular Biology of the Cell, Vol. 13, No. 11, 01.11.2002, p. 4001-4012.

Research output: Contribution to journalArticle

Yao, Diying ; Ehrlich, Marcelo ; Henis, Yoav I. ; Leof, Edward B. / Transforming growth factor-β receptors interact with AP2 by direct binding to β2 subunit. In: Molecular Biology of the Cell. 2002 ; Vol. 13, No. 11. pp. 4001-4012.
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