Cargando…

Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation

Irgb6 is a member of interferon γ-induced immunity related GTPase (IRG), and one of twenty “effector” IRGs, which coordinately attack parasitophorous vacuole membrane (PVM), causing death of intracellular pathogen. Although Irgb6 plays a pivotal role as a pioneer in the process of PVM disruption, th...

Descripción completa

Detalles Bibliográficos
Autores principales: Yamada, Hiroshi, Abe, Tadashi, Nagaoka, Hikaru, Takashima, Eizo, Nitta, Ryo, Yamamoto, Masahiro, Takei, Kohji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9504060/
https://www.ncbi.nlm.nih.gov/pubmed/36159643
http://dx.doi.org/10.3389/fcimb.2022.992198
_version_ 1784796121243058176
author Yamada, Hiroshi
Abe, Tadashi
Nagaoka, Hikaru
Takashima, Eizo
Nitta, Ryo
Yamamoto, Masahiro
Takei, Kohji
author_facet Yamada, Hiroshi
Abe, Tadashi
Nagaoka, Hikaru
Takashima, Eizo
Nitta, Ryo
Yamamoto, Masahiro
Takei, Kohji
author_sort Yamada, Hiroshi
collection PubMed
description Irgb6 is a member of interferon γ-induced immunity related GTPase (IRG), and one of twenty “effector” IRGs, which coordinately attack parasitophorous vacuole membrane (PVM), causing death of intracellular pathogen. Although Irgb6 plays a pivotal role as a pioneer in the process of PVM disruption, the direct effect of Irgb6 on membrane remained to be elucidated. Here, we utilized artificial lipid membranes to reconstitute Irgb6-membrane interaction in vitro, and revealed that Irgb6 directly deformed the membranes. Liposomes incubated with recombinant Irgb6 were drastically deformed generating massive tubular protrusions in the absence of guanine nucleotide, or with GMP-PNP. Liposome deformation was abolished by incubating with Irgb6-K275A/R371A, point mutations at membrane targeting residues. The membrane tubules generated by Irgb6 were mostly disappeared by the addition of GTP or GDP, which are caused by detachment of Irgb6 from membrane. Binding of Irgb6 to the membrane, which was reconstituted in vitro using lipid monolayer, was stimulated at GTP-bound state. Irgb6 GTPase activity was stimulated by the presence of liposomes more than eightfold. Irgb6 GTPase activity in the absence of membrane was also slightly stimulated, by lowering ionic strength, or by increasing protein concentration, indicating synergistic stimulation of the GTPase activity. These results suggest that membrane targeting of Irgb6 and resulting membrane deformation does not require GTP, but converting into GTP-bound state is crucial for detaching Irgb6 from the membrane, which might coincident with local membrane disruption.
format Online
Article
Text
id pubmed-9504060
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-95040602022-09-24 Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation Yamada, Hiroshi Abe, Tadashi Nagaoka, Hikaru Takashima, Eizo Nitta, Ryo Yamamoto, Masahiro Takei, Kohji Front Cell Infect Microbiol Cellular and Infection Microbiology Irgb6 is a member of interferon γ-induced immunity related GTPase (IRG), and one of twenty “effector” IRGs, which coordinately attack parasitophorous vacuole membrane (PVM), causing death of intracellular pathogen. Although Irgb6 plays a pivotal role as a pioneer in the process of PVM disruption, the direct effect of Irgb6 on membrane remained to be elucidated. Here, we utilized artificial lipid membranes to reconstitute Irgb6-membrane interaction in vitro, and revealed that Irgb6 directly deformed the membranes. Liposomes incubated with recombinant Irgb6 were drastically deformed generating massive tubular protrusions in the absence of guanine nucleotide, or with GMP-PNP. Liposome deformation was abolished by incubating with Irgb6-K275A/R371A, point mutations at membrane targeting residues. The membrane tubules generated by Irgb6 were mostly disappeared by the addition of GTP or GDP, which are caused by detachment of Irgb6 from membrane. Binding of Irgb6 to the membrane, which was reconstituted in vitro using lipid monolayer, was stimulated at GTP-bound state. Irgb6 GTPase activity was stimulated by the presence of liposomes more than eightfold. Irgb6 GTPase activity in the absence of membrane was also slightly stimulated, by lowering ionic strength, or by increasing protein concentration, indicating synergistic stimulation of the GTPase activity. These results suggest that membrane targeting of Irgb6 and resulting membrane deformation does not require GTP, but converting into GTP-bound state is crucial for detaching Irgb6 from the membrane, which might coincident with local membrane disruption. Frontiers Media S.A. 2022-09-09 /pmc/articles/PMC9504060/ /pubmed/36159643 http://dx.doi.org/10.3389/fcimb.2022.992198 Text en Copyright © 2022 Yamada, Abe, Nagaoka, Takashima, Nitta, Yamamoto and Takei https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cellular and Infection Microbiology
Yamada, Hiroshi
Abe, Tadashi
Nagaoka, Hikaru
Takashima, Eizo
Nitta, Ryo
Yamamoto, Masahiro
Takei, Kohji
Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation
title Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation
title_full Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation
title_fullStr Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation
title_full_unstemmed Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation
title_short Recruitment of Irgb6 to the membrane is a direct trigger for membrane deformation
title_sort recruitment of irgb6 to the membrane is a direct trigger for membrane deformation
topic Cellular and Infection Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9504060/
https://www.ncbi.nlm.nih.gov/pubmed/36159643
http://dx.doi.org/10.3389/fcimb.2022.992198
work_keys_str_mv AT yamadahiroshi recruitmentofirgb6tothemembraneisadirecttriggerformembranedeformation
AT abetadashi recruitmentofirgb6tothemembraneisadirecttriggerformembranedeformation
AT nagaokahikaru recruitmentofirgb6tothemembraneisadirecttriggerformembranedeformation
AT takashimaeizo recruitmentofirgb6tothemembraneisadirecttriggerformembranedeformation
AT nittaryo recruitmentofirgb6tothemembraneisadirecttriggerformembranedeformation
AT yamamotomasahiro recruitmentofirgb6tothemembraneisadirecttriggerformembranedeformation
AT takeikohji recruitmentofirgb6tothemembraneisadirecttriggerformembranedeformation