title: Calreticulin-dimerization induced by post-translational arginylation is critical for stress granules scaffolding.
abstract: Protein arginylation mediated by arginyl-tRNA protein transferase is a post-translational modification that occurs widely in biology, it has been shown to regulate protein and properties and functions. Post-translational arginylation is critical for embryogenesis, cardiovascular development and angiogenesis but the molecular effects of proteins arginylated in vivo are largely unknown. In the present study, we demonstrate that arginylation reduces CRT (calreticulin) thermostability and induces a greater degree of dimerization and oligomerization. R-CRT (arginylated calreticulin) forms disulfide-bridged dimers that are increased in low Ca(2+) conditions at physiological temperatures, a similar condition to the cellular environment that it required for arginylation of CRT. Moreover, R-CRT self-oligomerizes through non-covalent interactions that are enhanced at temperatures above 40 °C, condition that mimics the heat shock treatment where R-CRT is the only isoespecies of CRT that associates in cells to SGs (stress granules). We show that in cells lacking CRT the scaffolding of larger SGs is impaired; the transfection with CRT (hence R-CRT expression) restores SGs assembly whereas the transfection with CRT mutated in Cys146 does not. Thus, R-CRT disulfide-bridged dimers (through Cys146) are essential for the scaffolding of larger SGs under heat shock, although these dimers are not required for R-CRT association to SGs. The alteration in SGs assembly is critical for the normal cellular recover of cells after heat induced stress. We conclude that R-CRT is emerging as a novel protein that has an impact on the regulation of SGs scaffolding and cell survival.
authors.affiliation: Centro de Investigaciones en Química Biológica de Córdoba, CIQUIBIC, CONICET-Departamento de Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Haya de la Torre y Medina Allende X5000HUA, Córdoba, Argentina.
authors.forename: Marcos A
authors.initials: MA
authors.lastname: Carpio
authors.forename: María B
authors.initials: MB
authors.lastname: Decca
authors.forename: Cecilia
authors.initials: C
authors.lastname: Lopez Sambrooks
authors.forename: Edith S
authors.initials: ES
authors.lastname: Durand
authors.forename: Guillermo G
authors.initials: GG
authors.lastname: Montich
authors.forename: Marta E
authors.initials: ME
authors.lastname: Hallak
mesh_terms: D019881:Aminoacyltransferases; D000818:Animals; D017209:Apoptosis; D001120:Arginine / Q000737:chemistry*; D037282:Calreticulin / Q000737:chemistry* / Q000378:metabolism*; D002460:Cell Line; D003594:Cytoplasmic Granules / Q000378:metabolism; D019281:Dimerization; D006360:Heat-Shock Proteins / Q000378:metabolism*; D018869:Heat-Shock Response; D051379:Mice; D011499:Protein Processing, Post-Translational
chemical_list: D037282:Calreticulin; D006360:Heat-Shock Proteins; D001120:Arginine; D019881:Aminoacyltransferases; C019727:arginyltransferase