title: Affinity purification-mass spectrometry analysis of bcl-2 interactome identified SLIRP as a novel interacting protein.
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.
authors: Department of Research, Advanced Diagnostics and Technological Innovation, Regina Elena National Cancer Institute, Rome, Italy.
authors: D
authors: http://orcid.org/0000-0002-7007-7914
authors: D
authors: Trisciuoglio
authors: Department of Research, Advanced Diagnostics and Technological Innovation, Regina Elena National Cancer Institute, Rome, Italy.
authors: M
authors: M
authors: Desideri
authors: Department of Research, Advanced Diagnostics and Technological Innovation, Regina Elena National Cancer Institute, Rome, Italy.
authors: V
authors: V
authors: Farini
authors: Department of Research, Advanced Diagnostics and Technological Innovation, Regina Elena National Cancer Institute, Rome, Italy.
authors: T
authors: T
authors: De Luca
authors: Department of Research, Advanced Diagnostics and Technological Innovation, Regina Elena National Cancer Institute, Rome, Italy.
authors: M
authors: M
authors: Di Martile
authors: Department of Research, Advanced Diagnostics and Technological Innovation, Regina Elena National Cancer Institute, Rome, Italy.
authors: M G
authors: MG
authors: Tupone
authors: Department of Experimental Medicine and Surgery, University of 'Tor Vergata', Rome, Italy.
authors: A
authors: A
authors: Urbani
authors: Department of Research, Advanced Diagnostics and Technological Innovation, Regina Elena National Cancer Institute, Rome, Italy.
authors: S
authors: S
authors: D'Aguanno
authors: Department of Research, Advanced Diagnostics and Technological Innovation, Regina Elena National Cancer Institute, Rome, Italy.
authors: D
authors: D
authors: Del Bufalo
mesh_terms: D017209:Apoptosis / Q000502:physiology; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D019253:Proto-Oncogene Proteins c-bcl-2; D016601:RNA-Binding Proteins; D017382:Reactive Oxygen Species; C512286:SLIRP protein, human