title: New insights on the interaction between the isoforms 1 and 2 of human translation elongation factor 1A.
abstract: The eukaryotic translation elongation factor 1A (eEF1A) is a moonlighting protein that besides to its canonical role in protein synthesis is also involved in many other cellular processes such as cell survival and apoptosis. In a previous work, we identified eEF1A Raf-mediated phosphorylation sites and defined their role in the regulation of eEF1A half-life and apoptosis of human cancer cells. We proposed that the phosphorylation of eEF1A by C-Raf required the presence of both eEF1A isoforms thus suggesting the formation of a potential eEF1A heterodimer owning regulatory properties. This study aimed at investigating the cellular localization and interaction between two eEF1A isoforms. To this end, we developed chimera proteins by adding at the N-terminal end of both eEF1A1 and eEF1A2 cyan fluorescence protein (mCerulean) and yellow fluorescence protein (mVenus), respectively. The fluorescent eEF1A1 and eEF1A2 chimeras were both addressed to COS-7 cells and found co-localized in the cytoplasm at the level of cellular membranes. We highlighted FRET between the labeled N-termini of eEF1A isoforms. The intra-molecular FRET of this chimera was about 17%. Our results provide novel information on the intracellular distribution and interaction of eEF1A isoforms.
authors.affiliation: Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
authors.forename: Nunzia
authors.initials: N
authors.lastname: Migliaccio
authors.affiliation: Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
authors.forename: Immacolata
authors.initials: I
authors.lastname: Ruggiero
authors.affiliation: Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
authors.forename: Nicola M
authors.initials: NM
authors.lastname: Martucci
authors.affiliation: Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
authors.forename: Carmen
authors.initials: C
authors.lastname: Sanges
authors.affiliation: Institute of Genetics and Biophysics, Integrated Microscopy Facility, IGB CNR, Naples, Italy.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Arbucci
authors.affiliation: Institute of Genetics and Biophysics, Integrated Microscopy Facility, IGB CNR, Naples, Italy.
authors.forename: Rosarita
authors.initials: R
authors.lastname: Tatè
authors.affiliation: Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
authors.forename: Emilia
authors.initials: E
authors.lastname: Rippa
authors.affiliation: Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy; CEINGE, Advanced Biotechnology Scarl, Via Gaetano Salvatore 486, I-80145 Naples, Italy. Electronic address: paolo.arcari@unina.it.
authors.forename: Paolo
authors.initials: P
authors.lastname: Arcari
authors.affiliation: Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Lamberti
mesh_terms: D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002678:Chimera; D002522:Chlorocebus aethiops; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D018613:Microscopy, Confocal; D020648:Peptide Elongation Factor 1 / Q000378:metabolism*; D014162:Transfection
chemical_list: C094814:EEF1A1 protein, human; C503234:EEF1A2 protein, human; D020648:Peptide Elongation Factor 1
keywords: Confocal microscopy; Crosslinking; Eukaryotic translation elongation factor 1A (eEF1A); FRET