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Role of endocytosis in the activation of the extracellular signal-regulated kinase cascade by sequestering and nonsequestering G protein-coupled receptors.

Publication ,  Journal Article
Pierce, KL; Maudsley, S; Daaka, Y; Luttrell, LM; Lefkowitz, RJ
Published in: Proc Natl Acad Sci U S A
February 15, 2000

Acting through a number of distinct pathways, many G protein-coupled receptors (GPCRs) activate the extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase (MAPK) cascade. Recently, it has been shown that in some cases, clathrin-mediated endocytosis is required for GPCR activation of the ERK/MAPK cascade, whereas in others it is not. Accordingly, we compared ERK activation mediated by a GPCR that does not undergo agonist-stimulated endocytosis, the alpha(2A) adrenergic receptor (alpha(2A) AR), with ERK activation mediated by the beta(2) adrenergic receptor (beta(2) AR), which is endocytosed. Surprisingly, we found that in COS-7 cells, ERK activation by the alpha(2A) AR, like that mediated by both the beta(2) AR and the epidermal growth factor receptor (EGFR), is sensitive to mechanistically distinct inhibitors of clathrin-mediated endocytosis, including monodansylcadaverine, a mutant dynamin I, and a mutant beta-arrestin 1. Moreover, we determined that, as has been shown for many other GPCRs, both alpha(2A) and beta(2) AR-mediated ERK activation involves transactivation of the EGFR. Using confocal immunofluorescence microscopy, we found that stimulation of the beta(2) AR, the alpha(2A) AR, or the EGFR each results in internalization of a green fluorescent protein-tagged EGFR. Although beta(2) AR stimulation leads to redistribution of both the beta(2) AR and EGFR, activation of the alpha(2A) AR leads to redistribution of the EGFR but the alpha(2A) AR remains on the plasma membrane. These findings separate GPCR endocytosis from the requirement for clathrin-mediated endocytosis in EGFR transactivation-mediated ERK activation and suggest that it is the receptor tyrosine kinase or another downstream effector that must engage the endocytic machinery.

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Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

February 15, 2000

Volume

97

Issue

4

Start / End Page

1489 / 1494

Location

United States

Related Subject Headings

  • Tyrphostins
  • Transfection
  • Receptors, Cell Surface
  • Receptors, Adrenergic, beta
  • Receptors, Adrenergic, alpha
  • Quinoxalines
  • Quinazolines
  • Phosphorylation
  • Mitogen-Activated Protein Kinases
  • Isoproterenol
 

Citation

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Pierce, K. L., Maudsley, S., Daaka, Y., Luttrell, L. M., & Lefkowitz, R. J. (2000). Role of endocytosis in the activation of the extracellular signal-regulated kinase cascade by sequestering and nonsequestering G protein-coupled receptors. Proc Natl Acad Sci U S A, 97(4), 1489–1494. https://doi.org/10.1073/pnas.97.4.1489
Pierce, K. L., S. Maudsley, Y. Daaka, L. M. Luttrell, and R. J. Lefkowitz. “Role of endocytosis in the activation of the extracellular signal-regulated kinase cascade by sequestering and nonsequestering G protein-coupled receptors.Proc Natl Acad Sci U S A 97, no. 4 (February 15, 2000): 1489–94. https://doi.org/10.1073/pnas.97.4.1489.
Pierce KL, Maudsley S, Daaka Y, Luttrell LM, Lefkowitz RJ. Role of endocytosis in the activation of the extracellular signal-regulated kinase cascade by sequestering and nonsequestering G protein-coupled receptors. Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1489–94.
Pierce, K. L., et al. “Role of endocytosis in the activation of the extracellular signal-regulated kinase cascade by sequestering and nonsequestering G protein-coupled receptors.Proc Natl Acad Sci U S A, vol. 97, no. 4, Feb. 2000, pp. 1489–94. Pubmed, doi:10.1073/pnas.97.4.1489.
Pierce KL, Maudsley S, Daaka Y, Luttrell LM, Lefkowitz RJ. Role of endocytosis in the activation of the extracellular signal-regulated kinase cascade by sequestering and nonsequestering G protein-coupled receptors. Proc Natl Acad Sci U S A. 2000 Feb 15;97(4):1489–1494.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

February 15, 2000

Volume

97

Issue

4

Start / End Page

1489 / 1494

Location

United States

Related Subject Headings

  • Tyrphostins
  • Transfection
  • Receptors, Cell Surface
  • Receptors, Adrenergic, beta
  • Receptors, Adrenergic, alpha
  • Quinoxalines
  • Quinazolines
  • Phosphorylation
  • Mitogen-Activated Protein Kinases
  • Isoproterenol