Identification of FBXO25-interacting proteins using an integrated proteomics approach

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Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics
12:53

Identification of Protein Interaction Partners in Mammalian Cells Using SILAC-immunoprecipitation Quantitative Proteomics

Published on: July 6, 2014

Felipe R Teixeira1, Sami Yokoo, Carlos A Gartner

  • 1Department of Biochemistry and Immunology, University of São Paulo, Ribeirão Preto, Brazil.

Proteomics
|May 18, 2010
PubMed

Insights

FBXO25 protein forms novel nuclear structures called FAND, interacting with beta-actin. Actin polymerization influences these FAND compartments, impacting nuclear functions like transcription.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • FBXO25 is a human F-box protein involved in the SCF1 ubiquitin ligase complex.
  • SCF1 targets proteins for degradation via the ubiquitin-proteasome system.
  • FBXO25 has been observed to accumulate in novel subnuclear structures, termed FBXO25-associated nuclear domains (FAND).

Purpose of the Study:

  • To identify novel FBXO25 interacting partners.
  • To investigate the composition and regulation of FAND.
  • To explore the functional role of FBXO25 and FAND in nuclear processes.

Main Methods:

  • Two-step affinity purification followed by mass spectrometry (MS).
  • Classical two-hybrid screening.
  • In vitro transcription assays using FBXO25 antibodies.

Main Results:

  • Identified 132 novel potential FBXO25 interacting partners.
  • Beta-actin was identified as an FBXO25 interactor, localizing to FAND.
  • Actin polymerization inhibitors disrupted FAND, suggesting actin's role in their organization.
  • FBXO25 antibodies inhibited RNA polymerase II transcription in vitro.

Conclusions:

  • FBXO25 forms novel nuclear compartments (FAND) influenced by nuclear actin organization.
  • FBXO25 and FAND are implicated in regulating nuclear functions, including transcription.
  • These findings provide new insights into the function of FAND and FBXO25 in the nucleus.

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