Affinity purification-mass spectrometry analysis of bcl-2 interactome identified SLIRP as a novel interacting protein

D. Trisciuoglio, M. Desideri, V. Farini, T. De Luca, M. Di Martile, M. G. Tupone, A. Urbani, S. D'Aguanno, D. Del Bufalo

Research output: Contribution to journalArticle

Abstract

Members of the bcl-2 protein family share regions of sequence similarity, the bcl-2 homology (BH) domains. Bcl-2, the most studied member of this family, has four BH domains, BH1 - 4, and has a critical role in resistance to antineoplastic drugs by regulating the mitochondrial apoptotic pathway. Moreover, it is also involved in other relevant cellular processes such as tumor progression, angiogenesis and autophagy. Deciphering the network of bcl-2-interacting factors should provide a critical advance in understanding the different functions of bcl-2. Here, we characterized bcl-2 interactome by mass spectrometry in human lung adenocarcinoma cells. In silico functional analysis associated most part of the identified proteins to mitochondrial functions. Among them we identified SRA stem-loop interacting RNA-binding protein, SLIRP, a mitochondrial protein with a relevant role in regulating mitochondrial messenger RNA (mRNA) homeostasis. We validated bcl-2/SLIRP interaction by immunoprecipitation and immunofluorescence experiments in cancer cell lines from different histotypes. We showed that, although SLIRP is not involved in mediating bcl-2 ability to protect from apoptosis and oxidative damage, bcl-2 binds and stabilizes SLIRP protein and regulates mitochondrial mRNA levels. Moreover, we demonstrated that the BH4 domain of bcl-2 has a role in maintaining this binding.

Original languageEnglish
Article numbere2090
JournalCell Death and Disease
Volume7
Issue number2
DOIs
Publication statusPublished - 2016

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Mitochondrial Proteins
Neoplasm Drug Resistance
Mass Spectrometry
Aptitude
RNA-Binding Proteins
Autophagy
Immunoprecipitation
Computer Simulation
Fluorescent Antibody Technique
Neoplasms
Proteins
Homeostasis
Apoptosis
Cell Line
mitochondrial messenger RNA
Adenocarcinoma of lung

ASJC Scopus subject areas

  • Cell Biology
  • Immunology
  • Cancer Research
  • Cellular and Molecular Neuroscience

Cite this

Affinity purification-mass spectrometry analysis of bcl-2 interactome identified SLIRP as a novel interacting protein. / Trisciuoglio, D.; Desideri, M.; Farini, V.; De Luca, T.; Di Martile, M.; Tupone, M. G.; Urbani, A.; D'Aguanno, S.; Del Bufalo, D.

In: Cell Death and Disease, Vol. 7, No. 2, e2090, 2016.

Research output: Contribution to journalArticle

Trisciuoglio, D. ; Desideri, M. ; Farini, V. ; De Luca, T. ; Di Martile, M. ; Tupone, M. G. ; Urbani, A. ; D'Aguanno, S. ; Del Bufalo, D. / Affinity purification-mass spectrometry analysis of bcl-2 interactome identified SLIRP as a novel interacting protein. In: Cell Death and Disease. 2016 ; Vol. 7, No. 2.
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