ER membrane receptors of the GET pathway are conserved throughout eukaryotes

• Autores: Lisa Yasmin Asseck; Mehlhorn, Dietmar Gerald; Monroy, Jhon Rivera; Ricardi, Martiniano María; Breuninger, Holger; Wallmeroth, Niklas; Berendzen, Kenneth Wayne; Nowrousian, Minou; Xing, Shuping; Blanche Schwappach; Bayer, Martin; Grefen, Christopher
• Año de publicación: 2021
• Idioma: inglés
• Tipo de recurso: artículo
• Materia: ER MEMBRANE; GET PATHWAY; ROOT HAIRS; SNARES; TAIL-ANCHORED PROTEINS
• Institución: Consejo Nacional de Investigaciones Científicas y Técnicas

Type II tail-anchored (TA) membrane proteins are involved in diverse cellular processes, including protein translocation, vesicle trafficking, and apoptosis. They are characterized by a single C-terminal transmembrane domain that mediates posttranslational targeting and insertion into the endoplasmic reticulum (ER) via the Guided-Entry of TA proteins (GET) pathway. The GET system was originally described in mammals and yeast but was recently shown to be partially conserved in other eukaryotes, such as higher plants. A newly synthesized TA protein is shielded from the cytosol by a pretargeting complex and an ATPase that delivers the protein to the ER, where membrane receptors (Get1/WRB and Get2/CAML) facilitate insertion. In the model plant Arabidopsis thaliana, most components of the pathway were identified through in silico sequence comparison, however, a functional homolog of the coreceptor Get2/CAML remained elusive. We performed immunoprecipitation- mass spectrometry analysis to detect in vivo interactors of AtGET1 and identified a membrane protein of unknown function with low sequence homology but high structural homology to both yeast Get2 and mammalian CAML. The protein localizes to the ER membrane, coexpresses with AtGET1, and binds to Arabidopsis GET pathway components. While loss-of-function lines phenocopy the stunted root hair phenotype of other Atget lines, its heterologous expression together with the coreceptor AtGET1 rescues growth defects of Δget1get2 yeast. Ectopic expression of the cytosolic, positively charged N terminus is sufficient to block TA protein insertion in vitro. Our results collectively confirm that we have identified a plant-specific GET2 in Arabidopsis, and its sequence allows the analysis of cross-kingdom pathway conservation.
Fil: Lisa Yasmin Asseck. Eberhard Karls Universität Tübingen.; Alemania
Fil: Mehlhorn, Dietmar Gerald. Eberhard Karls Universität Tübingen.; Alemania
Fil: Monroy, Jhon Rivera. Universität Göttingen; Alemania
Fil: Ricardi, Martiniano María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina
Fil: Breuninger, Holger. Eberhard Karls Universität Tübingen.; Alemania
Fil: Wallmeroth, Niklas. Eberhard Karls Universität Tübingen.; Alemania
Fil: Berendzen, Kenneth Wayne. Eberhard Karls Universität Tübingen.; Alemania
Fil: Nowrousian, Minou. Ruhr Universität Bochum; Alemania
Fil: Xing, Shuping. Eberhard Karls Universität Tübingen.; Alemania
Fil: Blanche Schwappach. Universität Göttingen; Alemania
Fil: Bayer, Martin. Institut Max Planck Fuer Gesellschaft. Institut Fur Entwicklungsbiolobie. Developmental Biology; Alemania
Fil: Grefen, Christopher. Eberhard Karls Universität Tübingen.; Alemania