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Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes

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 endoplasmi...

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Autores principales: Asseck, Lisa Yasmin, Mehlhorn, Dietmar Gerald, Monroy, Jhon Rivera, Ricardi, Martiniano Maria, Breuninger, Holger, Wallmeroth, Niklas, Berendzen, Kenneth Wayne, Nowrousian, Minou, Xing, Shuping, Schwappach, Blanche, Bayer, Martin, Grefen, Christopher
Formato: Online Artículo Texto
Lenguaje:English
Publicado: National Academy of Sciences 2021
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7817167/
https://www.ncbi.nlm.nih.gov/pubmed/33443185
http://dx.doi.org/10.1073/pnas.2017636118
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author Asseck, Lisa Yasmin
Mehlhorn, Dietmar Gerald
Monroy, Jhon Rivera
Ricardi, Martiniano Maria
Breuninger, Holger
Wallmeroth, Niklas
Berendzen, Kenneth Wayne
Nowrousian, Minou
Xing, Shuping
Schwappach, Blanche
Bayer, Martin
Grefen, Christopher
author_facet Asseck, Lisa Yasmin
Mehlhorn, Dietmar Gerald
Monroy, Jhon Rivera
Ricardi, Martiniano Maria
Breuninger, Holger
Wallmeroth, Niklas
Berendzen, Kenneth Wayne
Nowrousian, Minou
Xing, Shuping
Schwappach, Blanche
Bayer, Martin
Grefen, Christopher
author_sort Asseck, Lisa Yasmin
collection PubMed
description 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.
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spelling pubmed-78171672021-01-28 Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes Asseck, Lisa Yasmin Mehlhorn, Dietmar Gerald Monroy, Jhon Rivera Ricardi, Martiniano Maria Breuninger, Holger Wallmeroth, Niklas Berendzen, Kenneth Wayne Nowrousian, Minou Xing, Shuping Schwappach, Blanche Bayer, Martin Grefen, Christopher Proc Natl Acad Sci U S A Biological Sciences 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. National Academy of Sciences 2021-01-05 2020-12-28 /pmc/articles/PMC7817167/ /pubmed/33443185 http://dx.doi.org/10.1073/pnas.2017636118 Text en Copyright © 2021 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/ https://creativecommons.org/licenses/by-nc-nd/4.0/This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Biological Sciences
Asseck, Lisa Yasmin
Mehlhorn, Dietmar Gerald
Monroy, Jhon Rivera
Ricardi, Martiniano Maria
Breuninger, Holger
Wallmeroth, Niklas
Berendzen, Kenneth Wayne
Nowrousian, Minou
Xing, Shuping
Schwappach, Blanche
Bayer, Martin
Grefen, Christopher
Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes
title Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes
title_full Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes
title_fullStr Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes
title_full_unstemmed Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes
title_short Endoplasmic reticulum membrane receptors of the GET pathway are conserved throughout eukaryotes
title_sort endoplasmic reticulum membrane receptors of the get pathway are conserved throughout eukaryotes
topic Biological Sciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7817167/
https://www.ncbi.nlm.nih.gov/pubmed/33443185
http://dx.doi.org/10.1073/pnas.2017636118
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