IHEM   20887
INSTITUTO DE HISTOLOGIA Y EMBRIOLOGIA DE MENDOZA DR. MARIO H. BURGOS
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
14-3-3-Prohibitin interactions. In vivo and in silico studies.
Autor/es:
MARINA UHART; DESAUBRY; DIEGO M. BUSTOS
Lugar:
Rosario
Reunión:
Congreso; L Reunión Anual de la Sociedad Argentina de Investigaciones Bioquímicas y Biología Molecular (SAIB); 2014
Institución organizadora:
SAIB
Resumen:
The 14-3-3 protein family is a phosphoproteome master regulator. It binds more than 2000 cellular proteins, recognizing two structural features in its client proteins: a serine or threonine phospho residue and a region of about 30 consecutive amino acids rich in residues that promote intrinsic structural disorder. Although 14-3-3 is described as a ubiquitous protein family, most of its complexes found in the literature include either cytosolic or nuclear proteins. Here, we show the interaction of 14-3-3 with Prohibitin 1 and 2 (PHB1 and 2), a mitochondrial and fully structured protein, therefore an atypical 14-3-3 partner. Both, 14-3-3s and PHBs, are hubs for many signaling pathways triggered by growth factors, the immune response, and steroid hormones regulating metabolism, mitochondrial biogenesis, cell migration, division, and survival. We used in vivo Föster Resonance Energy Transfer (FRET) of genetically encoded fluorescent CFP-14-3-3 beta and PHBs-YFP to demonstrate the interaction between these two proteins and to show the subcellular localization of the complex. Transfected HeLa cells showed positive FRET signal specifically localized in the mitochondria, and no signal was observed when a nuclear 14-3-3 (CFP-14-3-3-NLS) was used as control. A possible structure of the complex is proposed by in silico molecular docking using the structures of both proteins from the PDB (1LU7 and 1A40).