Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes
| العنوان: | Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes |
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| المؤلفون: | Martiniano Maria Ricardi, Holger Breuninger, Lisa Yasmin Asseck, Martin Bayer, Christopher Grefen, Niklas Wallmeroth, Minou Nowrousian, Shuping Xing, Dietmar G. Mehlhorn, Kenneth W. Berendzen, Blanche Schwappach, Jhon Rivera Monroy |
| المصدر: | Proceedings of the National Academy of Sciences of the United States of America |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | 0106 biological sciences, SNAREs, Saccharomyces cerevisiae Proteins, ER membrane, Arabidopsis, Plant Biology, Saccharomyces cerevisiae, GET pathway, Endoplasmic Reticulum, 01 natural sciences, root hairs, 03 medical and health sciences, Cytosol, Arabidopsis thaliana, tail-anchored proteins, 030304 developmental biology, computer.programming_language, 0303 health sciences, Multidisciplinary, Endoplasmic reticulum membrane, biology, Caml, Chemistry, Arabidopsis Proteins, Endoplasmic reticulum, Cell Membrane, Membrane Proteins, Intracellular Membranes, Biological Sciences, biology.organism_classification, Cell biology, Transmembrane domain, Adaptor Proteins, Vesicular Transport, Protein Transport, Phenotype, Membrane protein, Heterologous expression, computer, 010606 plant biology & botany |
| الوصف: | Significance The GET pathway is required for the insertion of tail-anchored (TA) membrane proteins in the endoplasmic reticulum (ER) of yeast and mammals. Some orthologous genes had also been identified in higher plants with the exception of one of the two ER membrane receptors required for membrane insertion. Get2/CAML is required for the pathway’s cytosolic chaperone to dock and release its TA protein cargo. Here we report the identification of the elusive plant GET pathway receptor through an interaction screen in Arabidopsis. The candidate allows detection of further Get2/CAML orthologs in higher plants, revealing conservation and function of structural features across kingdoms. Additionally, our results demonstrate that these features, rather than sequence conservation, determine functionality of the candidate within the pathway. 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. |
| تدمد: | 1091-6490 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4316cffd90f2e2ffd8fb9cf048d28407 https://pubmed.ncbi.nlm.nih.gov/33443185 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....4316cffd90f2e2ffd8fb9cf048d28407 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 10916490 |
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