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The Legionella longbeachae Icm/Dot substrate SidC selectively binds phosphatidylinositol 4-phosphate with nanomolar affinity and promotes pathogen vacuole-endoplasmic reticulum interactions

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The Legionella longbeachae Icm/Dot substrate SidC selectively binds phosphatidylinositol 4-phosphate with nanomolar affinity and promotes pathogen vacuole-endoplasmic reticulum interactions. / Dolinsky, Stephanie; Haneburger, Ina; Cichy, Adam; Hannemann, Mandy; Itzen, Aymelt; Hilbi, Hubert.

in: INFECT IMMUN, Jahrgang 82, Nr. 10, 10.2014, S. 4021-33.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Dolinsky, S, Haneburger, I, Cichy, A, Hannemann, M, Itzen, A & Hilbi, H 2014, 'The Legionella longbeachae Icm/Dot substrate SidC selectively binds phosphatidylinositol 4-phosphate with nanomolar affinity and promotes pathogen vacuole-endoplasmic reticulum interactions', INFECT IMMUN, Jg. 82, Nr. 10, S. 4021-33. https://doi.org/10.1128/IAI.01685-14

APA

Dolinsky, S., Haneburger, I., Cichy, A., Hannemann, M., Itzen, A., & Hilbi, H. (2014). The Legionella longbeachae Icm/Dot substrate SidC selectively binds phosphatidylinositol 4-phosphate with nanomolar affinity and promotes pathogen vacuole-endoplasmic reticulum interactions. INFECT IMMUN, 82(10), 4021-33. https://doi.org/10.1128/IAI.01685-14

Vancouver

Dolinsky S, Haneburger I, Cichy A, Hannemann M, Itzen A, Hilbi H. The Legionella longbeachae Icm/Dot substrate SidC selectively binds phosphatidylinositol 4-phosphate with nanomolar affinity and promotes pathogen vacuole-endoplasmic reticulum interactions. INFECT IMMUN. 2014 Okt;82(10):4021-33. https://doi.org/10.1128/IAI.01685-14

Bibtex

@article{246a2d8e49de44dfa12c74f699386082,
title = "The Legionella longbeachae Icm/Dot substrate SidC selectively binds phosphatidylinositol 4-phosphate with nanomolar affinity and promotes pathogen vacuole-endoplasmic reticulum interactions",
abstract = "Legionella spp. cause the severe pneumonia Legionnaires' disease. The environmental bacteria replicate intracellularly in free-living amoebae and human alveolar macrophages within a distinct, endoplasmic reticulum (ER)-derived compartment termed the Legionella-containing vacuole (LCV). LCV formation requires the bacterial Icm/Dot type IV secretion system (T4SS) that translocates into host cells a plethora of different {"}effector{"} proteins, some of which anchor to the pathogen vacuole by binding to phosphoinositide (PI) lipids. Here, we identified by unbiased pulldown assays in Legionella longbeachae lysates a 111-kDa SidC homologue as the major phosphatidylinositol 4-phosphate [PtdIns(4)P]-binding protein. The PI-binding domain was mapped to a 20-kDa P4C [PtdIns(4)P binding of SidC] fragment. Isothermal titration calorimetry revealed that SidC of L. longbeachae (SidC(Llo)) binds PtdIns(4)P with a K(d) (dissociation constant) of 71 nM, which is 3 to 4 times lower than that of the SidC orthologue of Legionella pneumophila (SidC(Lpn)). Upon infection of RAW 264.7 macrophages with L. longbeachae, endogenous SidC(Llo) or ectopically produced SidC(Lpn) localized in an Icm/Dot-dependent manner to the PtdIns(4)P-positive LCVs. An L. longbeachae ΔsidC deletion mutant was impaired for calnexin recruitment to LCVs in Dictyostelium discoideum amoebae and outcompeted by wild-type bacteria in Acanthamoeba castellanii. Calnexin recruitment was restored by SidC(Llo) or its orthologues SidC(Lpn) and SdcA(Lpn). Conversely, calnexin recruitment was restored by SidC(Llo) in L. pneumophila lacking sidC and sdcA. Together, biochemical, genetic, and cell biological data indicate that SidC(Llo) is an L. longbeachae effector that binds through a P4C domain with high affinity to PtdIns(4)P on LCVs, promotes ER recruitment to the LCV, and thus plays a role in pathogen-host interactions.",
keywords = "Acanthamoeba castellanii, Animals, Bacterial Proteins, Calnexin, Calorimetry, Cell Line, Chromosome Mapping, Dictyostelium, Endoplasmic Reticulum, Gene Deletion, Host-Pathogen Interactions, Kinetics, Legionella longbeachae, Macrophages, Mice, Molecular Weight, Phosphatidylinositol Phosphates, Protein Binding, Vacuoles, Journal Article, Research Support, Non-U.S. Gov't",
author = "Stephanie Dolinsky and Ina Haneburger and Adam Cichy and Mandy Hannemann and Aymelt Itzen and Hubert Hilbi",
note = "Copyright {\textcopyright} 2014, American Society for Microbiology. All Rights Reserved.",
year = "2014",
month = oct,
doi = "10.1128/IAI.01685-14",
language = "English",
volume = "82",
pages = "4021--33",
journal = "INFECT IMMUN",
issn = "0019-9567",
publisher = "American Society for Microbiology",
number = "10",

}

RIS

TY - JOUR

T1 - The Legionella longbeachae Icm/Dot substrate SidC selectively binds phosphatidylinositol 4-phosphate with nanomolar affinity and promotes pathogen vacuole-endoplasmic reticulum interactions

AU - Dolinsky, Stephanie

AU - Haneburger, Ina

AU - Cichy, Adam

AU - Hannemann, Mandy

AU - Itzen, Aymelt

AU - Hilbi, Hubert

N1 - Copyright © 2014, American Society for Microbiology. All Rights Reserved.

PY - 2014/10

Y1 - 2014/10

N2 - Legionella spp. cause the severe pneumonia Legionnaires' disease. The environmental bacteria replicate intracellularly in free-living amoebae and human alveolar macrophages within a distinct, endoplasmic reticulum (ER)-derived compartment termed the Legionella-containing vacuole (LCV). LCV formation requires the bacterial Icm/Dot type IV secretion system (T4SS) that translocates into host cells a plethora of different "effector" proteins, some of which anchor to the pathogen vacuole by binding to phosphoinositide (PI) lipids. Here, we identified by unbiased pulldown assays in Legionella longbeachae lysates a 111-kDa SidC homologue as the major phosphatidylinositol 4-phosphate [PtdIns(4)P]-binding protein. The PI-binding domain was mapped to a 20-kDa P4C [PtdIns(4)P binding of SidC] fragment. Isothermal titration calorimetry revealed that SidC of L. longbeachae (SidC(Llo)) binds PtdIns(4)P with a K(d) (dissociation constant) of 71 nM, which is 3 to 4 times lower than that of the SidC orthologue of Legionella pneumophila (SidC(Lpn)). Upon infection of RAW 264.7 macrophages with L. longbeachae, endogenous SidC(Llo) or ectopically produced SidC(Lpn) localized in an Icm/Dot-dependent manner to the PtdIns(4)P-positive LCVs. An L. longbeachae ΔsidC deletion mutant was impaired for calnexin recruitment to LCVs in Dictyostelium discoideum amoebae and outcompeted by wild-type bacteria in Acanthamoeba castellanii. Calnexin recruitment was restored by SidC(Llo) or its orthologues SidC(Lpn) and SdcA(Lpn). Conversely, calnexin recruitment was restored by SidC(Llo) in L. pneumophila lacking sidC and sdcA. Together, biochemical, genetic, and cell biological data indicate that SidC(Llo) is an L. longbeachae effector that binds through a P4C domain with high affinity to PtdIns(4)P on LCVs, promotes ER recruitment to the LCV, and thus plays a role in pathogen-host interactions.

AB - Legionella spp. cause the severe pneumonia Legionnaires' disease. The environmental bacteria replicate intracellularly in free-living amoebae and human alveolar macrophages within a distinct, endoplasmic reticulum (ER)-derived compartment termed the Legionella-containing vacuole (LCV). LCV formation requires the bacterial Icm/Dot type IV secretion system (T4SS) that translocates into host cells a plethora of different "effector" proteins, some of which anchor to the pathogen vacuole by binding to phosphoinositide (PI) lipids. Here, we identified by unbiased pulldown assays in Legionella longbeachae lysates a 111-kDa SidC homologue as the major phosphatidylinositol 4-phosphate [PtdIns(4)P]-binding protein. The PI-binding domain was mapped to a 20-kDa P4C [PtdIns(4)P binding of SidC] fragment. Isothermal titration calorimetry revealed that SidC of L. longbeachae (SidC(Llo)) binds PtdIns(4)P with a K(d) (dissociation constant) of 71 nM, which is 3 to 4 times lower than that of the SidC orthologue of Legionella pneumophila (SidC(Lpn)). Upon infection of RAW 264.7 macrophages with L. longbeachae, endogenous SidC(Llo) or ectopically produced SidC(Lpn) localized in an Icm/Dot-dependent manner to the PtdIns(4)P-positive LCVs. An L. longbeachae ΔsidC deletion mutant was impaired for calnexin recruitment to LCVs in Dictyostelium discoideum amoebae and outcompeted by wild-type bacteria in Acanthamoeba castellanii. Calnexin recruitment was restored by SidC(Llo) or its orthologues SidC(Lpn) and SdcA(Lpn). Conversely, calnexin recruitment was restored by SidC(Llo) in L. pneumophila lacking sidC and sdcA. Together, biochemical, genetic, and cell biological data indicate that SidC(Llo) is an L. longbeachae effector that binds through a P4C domain with high affinity to PtdIns(4)P on LCVs, promotes ER recruitment to the LCV, and thus plays a role in pathogen-host interactions.

KW - Acanthamoeba castellanii

KW - Animals

KW - Bacterial Proteins

KW - Calnexin

KW - Calorimetry

KW - Cell Line

KW - Chromosome Mapping

KW - Dictyostelium

KW - Endoplasmic Reticulum

KW - Gene Deletion

KW - Host-Pathogen Interactions

KW - Kinetics

KW - Legionella longbeachae

KW - Macrophages

KW - Mice

KW - Molecular Weight

KW - Phosphatidylinositol Phosphates

KW - Protein Binding

KW - Vacuoles

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1128/IAI.01685-14

DO - 10.1128/IAI.01685-14

M3 - SCORING: Journal article

C2 - 25024371

VL - 82

SP - 4021

EP - 4033

JO - INFECT IMMUN

JF - INFECT IMMUN

SN - 0019-9567

IS - 10

ER -