Desensitization, internalization, and signaling functions of beta-arrestins demonstrated by RNA interference

Proc Natl Acad Sci U S A. 2003 Feb 18;100(4):1740-4. doi: 10.1073/pnas.262789099. Epub 2003 Feb 11.

Abstract

Beta-arrestins bind to activated G protein-coupled receptor kinase-phosphorylated receptors, which leads to their desensitization with respect to G proteins, internalization via clathrin-coated pits, and signaling via a growing list of "scaffolded" pathways. To facilitate the discovery of novel adaptor and signaling roles of beta-arrestins, we have developed and validated a generally applicable interfering RNA approach for selectively suppressing beta-arrestins 1 or 2 expression by up to 95%. Beta-arrestin depletion in HEK293 cells leads to enhanced cAMP generation in response to beta(2)-adrenergic receptor stimulation, markedly reduced beta(2)-adrenergic receptor and angiotensin II receptor internalization and impaired activation of the MAP kinases ERK 1 and 2 by angiotensin II. This approach should allow discovery of novel signaling and regulatory roles for the beta-arrestins in many seven-membrane-spanning receptor systems.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Arrestins / metabolism*
  • Base Sequence
  • Blotting, Western
  • Cell Line
  • DNA Primers
  • Endocytosis*
  • Enzyme Activation
  • Humans
  • Mitogen-Activated Protein Kinase 1 / metabolism
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases / metabolism
  • Molecular Sequence Data
  • Phosphorylation
  • RNA, Small Interfering / genetics*
  • Signal Transduction*
  • beta-Arrestins

Substances

  • Arrestins
  • DNA Primers
  • RNA, Small Interfering
  • beta-Arrestins
  • Mitogen-Activated Protein Kinase 1
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinases

Associated data

  • RefSeq/NM_004313
  • RefSeq/NM_020251