Structural insight into the membrane targeting domain of the Legionella deAMPylase SidD

AMPylation, the post-translational modification with adenosine monophosphate (AMP), is catalyzed by effector proteins from a variety of pathogens. Legionella pneumophila is thus far the only known pathogen that, in addition to encoding an AMPylase (SidM/DrrA), also encodes a deAMPylase, called SidD,...

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Authors: Tascón, Igor, Li, Xiao, Lucas Gay, María|||0000-0002-7854-4249, Delson, D'anna, Vidaurrazaga, Ander, Lin, Yi-Han, Rojas, Adriana L., Hierro, Aitor, Machner, Matthias P
Format: article
Publication Date:2020
Country:España
Institution:Universidad de Cantabria (UC)
Repository:UCrea Repositorio Abierto de la Universidad de Cantabria
Language:English
OAI Identifier:oai:repositorio.unican.es:10902/20615
Online Access:http://hdl.handle.net/10902/20615
Access Level:Open access
Keyword:Adenosine Monophosphate
Legionella pneumophila
Cell Membrane
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spelling Structural insight into the membrane targeting domain of the Legionella deAMPylase SidDTascón, IgorLi, XiaoLucas Gay, María|||0000-0002-7854-4249Delson, D'annaVidaurrazaga, AnderLin, Yi-HanRojas, Adriana L.Hierro, AitorMachner, Matthias PAdenosine MonophosphateLegionella pneumophilaCell MembraneAMPylation, the post-translational modification with adenosine monophosphate (AMP), is catalyzed by effector proteins from a variety of pathogens. Legionella pneumophila is thus far the only known pathogen that, in addition to encoding an AMPylase (SidM/DrrA), also encodes a deAMPylase, called SidD, that reverses SidM-mediated AMPylation of the vesicle transport GTPase Rab1. DeAMPylation is catalyzed by the N-terminal phosphatase-like domain of SidD. Here, we determined the crystal structure of full length SidD including the uncharacterized C-terminal domain (CTD). A flexible loop rich in aromatic residues within the CTD was required to target SidD to model membranes in vitro and to the Golgi apparatus within mammalian cells. Deletion of the loop (??loop) or substitution of its aromatic phenylalanine residues rendered SidD cytosolic, showing that the hydrophobic loop is the primary membrane-targeting determinant of SidD. Notably, deletion of the two terminal alpha helices resulted in a CTD variant incapable of discriminating between membranes of different composition. Moreover, a L. pneumophila strain producing SidD??loop phenocopied a L. pneumophila ??sidD strain during growth in mouse macrophages and displayed prolonged co-localization of AMPylated Rab1 with LCVs, thus revealing that membrane targeting of SidD via its CTD is a critical prerequisite for its ability to catalyze Rab1 deAMPylation during L. pneumophila infection.Public Library of ScienceUniversidad de Cantabria20202020-01-01journal articlehttp://purl.org/coar/resource_type/c_6501NAhttp://purl.org/coar/version/c_be7fb7dd8ff6fe43info:eu-repo/semantics/articlehttp://hdl.handle.net/10902/20615PLoS Pathog. 2020 Aug; 16(8): e1008734reponame:UCrea Repositorio Abierto de la Universidad de Cantabriainstname:Universidad de Cantabria (UC)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:repositorio.unican.es:10902/206152026-06-02T12:39:31Z
dc.title.none.fl_str_mv Structural insight into the membrane targeting domain of the Legionella deAMPylase SidD
title Structural insight into the membrane targeting domain of the Legionella deAMPylase SidD
spellingShingle Structural insight into the membrane targeting domain of the Legionella deAMPylase SidD
Tascón, Igor
Adenosine Monophosphate
Legionella pneumophila
Cell Membrane
title_short Structural insight into the membrane targeting domain of the Legionella deAMPylase SidD
title_full Structural insight into the membrane targeting domain of the Legionella deAMPylase SidD
title_fullStr Structural insight into the membrane targeting domain of the Legionella deAMPylase SidD
title_full_unstemmed Structural insight into the membrane targeting domain of the Legionella deAMPylase SidD
title_sort Structural insight into the membrane targeting domain of the Legionella deAMPylase SidD
dc.creator.none.fl_str_mv Tascón, Igor
Li, Xiao
Lucas Gay, María|||0000-0002-7854-4249
Delson, D'anna
Vidaurrazaga, Ander
Lin, Yi-Han
Rojas, Adriana L.
Hierro, Aitor
Machner, Matthias P
author Tascón, Igor
author_facet Tascón, Igor
Li, Xiao
Lucas Gay, María|||0000-0002-7854-4249
Delson, D'anna
Vidaurrazaga, Ander
Lin, Yi-Han
Rojas, Adriana L.
Hierro, Aitor
Machner, Matthias P
author_role author
author2 Li, Xiao
Lucas Gay, María|||0000-0002-7854-4249
Delson, D'anna
Vidaurrazaga, Ander
Lin, Yi-Han
Rojas, Adriana L.
Hierro, Aitor
Machner, Matthias P
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidad de Cantabria
dc.subject.none.fl_str_mv Adenosine Monophosphate
Legionella pneumophila
Cell Membrane
topic Adenosine Monophosphate
Legionella pneumophila
Cell Membrane
description AMPylation, the post-translational modification with adenosine monophosphate (AMP), is catalyzed by effector proteins from a variety of pathogens. Legionella pneumophila is thus far the only known pathogen that, in addition to encoding an AMPylase (SidM/DrrA), also encodes a deAMPylase, called SidD, that reverses SidM-mediated AMPylation of the vesicle transport GTPase Rab1. DeAMPylation is catalyzed by the N-terminal phosphatase-like domain of SidD. Here, we determined the crystal structure of full length SidD including the uncharacterized C-terminal domain (CTD). A flexible loop rich in aromatic residues within the CTD was required to target SidD to model membranes in vitro and to the Golgi apparatus within mammalian cells. Deletion of the loop (??loop) or substitution of its aromatic phenylalanine residues rendered SidD cytosolic, showing that the hydrophobic loop is the primary membrane-targeting determinant of SidD. Notably, deletion of the two terminal alpha helices resulted in a CTD variant incapable of discriminating between membranes of different composition. Moreover, a L. pneumophila strain producing SidD??loop phenocopied a L. pneumophila ??sidD strain during growth in mouse macrophages and displayed prolonged co-localization of AMPylated Rab1 with LCVs, thus revealing that membrane targeting of SidD via its CTD is a critical prerequisite for its ability to catalyze Rab1 deAMPylation during L. pneumophila infection.
publishDate 2020
dc.date.none.fl_str_mv 2020
2020-01-01
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
NA
http://purl.org/coar/version/c_be7fb7dd8ff6fe43
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10902/20615
url http://hdl.handle.net/10902/20615
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv PLoS Pathog. 2020 Aug; 16(8): e1008734
reponame:UCrea Repositorio Abierto de la Universidad de Cantabria
instname:Universidad de Cantabria (UC)
instname_str Universidad de Cantabria (UC)
reponame_str UCrea Repositorio Abierto de la Universidad de Cantabria
collection UCrea Repositorio Abierto de la Universidad de Cantabria
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,300719

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