Legionella effector AnkX interacts with host nuclear protein PLEKHN1

Xiaobo Yu, Rebecca R. Noll, Barbara P. Romero Dueñas, Samual C. Allgood, Kristi Barker, Jeffrey L. Caplan, Matthias P. Machner, Joshua LaBaer, Ji Qiu, M. Ramona Neunuebel

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: The intracellular bacterial pathogen Legionella pneumophila proliferates in human alveolar macrophages, resulting in a severe pneumonia termed Legionnaires' disease. Throughout the course of infection, L. pneumophila remains enclosed in a specialized membrane compartment that evades fusion with lysosomes. The pathogen delivers over 300 effector proteins into the host cell, altering host pathways in a manner that sets the stage for efficient pathogen replication. The L. pneumophila effector protein AnkX targets host Rab GTPases and functions in preventing fusion of the Legionella-containing vacuole with lysosomes. However, the current understanding of AnkX's interaction with host proteins and the means through which it exerts its cellular function is limited. Results: Here, we investigated the protein interaction network of AnkX by using the nucleic acid programmable protein array (NAPPA), a high-density platform comprising 10,000 unique human ORFs. This approach facilitated the discovery of PLEKHN1 as a novel interaction partner of AnkX. We confirmed this interaction through multiple independent in vitro pull-down, co-immunoprecipitation, and cell-based assays. Structured illumination microscopy revealed that endogenous PLEKHN1 is found in the nucleus and on vesicular compartments, whereas ectopically produced AnkX co-localized with lipid rafts at the plasma membrane. In mammalian cells, HaloTag-AnkX co-localized with endogenous PLEKHN1 on vesicular compartments. A central fragment of AnkX (amino acids 491-809), containing eight ankyrin repeats, extensively co-localized with endogenous PLEKHN1, indicating that this region may harbor a new function. Further, we found that PLEKHN1 associated with multiple proteins involved in the inflammatory response. Conclusions: Altogether, our study provides evidence that in addition to Rab GTPases, the L. pneumophila effector AnkX targets nuclear host proteins and suggests that AnkX may have novel functions related to manipulating the inflammatory response.

Original languageEnglish (US)
Article number5
JournalBMC Microbiology
Volume18
Issue number1
DOIs
StatePublished - Jan 5 2018

Fingerprint

Legionella
Nuclear Proteins
Legionella pneumophila
rab GTP-Binding Proteins
Legionnaires' Disease
Lysosomes
Proteins
Ankyrin Repeat
Protein Interaction Maps
Protein Array Analysis
Alveolar Macrophages
Vacuoles
Lighting
Immunoprecipitation
Nucleic Acids
Open Reading Frames
Microscopy
Pneumonia
Cell Membrane
Lipids

Keywords

  • AnkX
  • Nucleic acid programmable protein array
  • PLEKHN1

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

Cite this

Yu, X., Noll, R. R., Romero Dueñas, B. P., Allgood, S. C., Barker, K., Caplan, J. L., ... Neunuebel, M. R. (2018). Legionella effector AnkX interacts with host nuclear protein PLEKHN1. BMC Microbiology, 18(1), [5]. https://doi.org/10.1186/s12866-017-1147-7

Legionella effector AnkX interacts with host nuclear protein PLEKHN1. / Yu, Xiaobo; Noll, Rebecca R.; Romero Dueñas, Barbara P.; Allgood, Samual C.; Barker, Kristi; Caplan, Jeffrey L.; Machner, Matthias P.; LaBaer, Joshua; Qiu, Ji; Neunuebel, M. Ramona.

In: BMC Microbiology, Vol. 18, No. 1, 5, 05.01.2018.

Research output: Contribution to journalArticle

Yu, X, Noll, RR, Romero Dueñas, BP, Allgood, SC, Barker, K, Caplan, JL, Machner, MP, LaBaer, J, Qiu, J & Neunuebel, MR 2018, 'Legionella effector AnkX interacts with host nuclear protein PLEKHN1', BMC Microbiology, vol. 18, no. 1, 5. https://doi.org/10.1186/s12866-017-1147-7
Yu X, Noll RR, Romero Dueñas BP, Allgood SC, Barker K, Caplan JL et al. Legionella effector AnkX interacts with host nuclear protein PLEKHN1. BMC Microbiology. 2018 Jan 5;18(1). 5. https://doi.org/10.1186/s12866-017-1147-7
Yu, Xiaobo ; Noll, Rebecca R. ; Romero Dueñas, Barbara P. ; Allgood, Samual C. ; Barker, Kristi ; Caplan, Jeffrey L. ; Machner, Matthias P. ; LaBaer, Joshua ; Qiu, Ji ; Neunuebel, M. Ramona. / Legionella effector AnkX interacts with host nuclear protein PLEKHN1. In: BMC Microbiology. 2018 ; Vol. 18, No. 1.
@article{2e35b8e0eea54f338d01e2b22c9fcd32,
title = "Legionella effector AnkX interacts with host nuclear protein PLEKHN1",
abstract = "Background: The intracellular bacterial pathogen Legionella pneumophila proliferates in human alveolar macrophages, resulting in a severe pneumonia termed Legionnaires' disease. Throughout the course of infection, L. pneumophila remains enclosed in a specialized membrane compartment that evades fusion with lysosomes. The pathogen delivers over 300 effector proteins into the host cell, altering host pathways in a manner that sets the stage for efficient pathogen replication. The L. pneumophila effector protein AnkX targets host Rab GTPases and functions in preventing fusion of the Legionella-containing vacuole with lysosomes. However, the current understanding of AnkX's interaction with host proteins and the means through which it exerts its cellular function is limited. Results: Here, we investigated the protein interaction network of AnkX by using the nucleic acid programmable protein array (NAPPA), a high-density platform comprising 10,000 unique human ORFs. This approach facilitated the discovery of PLEKHN1 as a novel interaction partner of AnkX. We confirmed this interaction through multiple independent in vitro pull-down, co-immunoprecipitation, and cell-based assays. Structured illumination microscopy revealed that endogenous PLEKHN1 is found in the nucleus and on vesicular compartments, whereas ectopically produced AnkX co-localized with lipid rafts at the plasma membrane. In mammalian cells, HaloTag-AnkX co-localized with endogenous PLEKHN1 on vesicular compartments. A central fragment of AnkX (amino acids 491-809), containing eight ankyrin repeats, extensively co-localized with endogenous PLEKHN1, indicating that this region may harbor a new function. Further, we found that PLEKHN1 associated with multiple proteins involved in the inflammatory response. Conclusions: Altogether, our study provides evidence that in addition to Rab GTPases, the L. pneumophila effector AnkX targets nuclear host proteins and suggests that AnkX may have novel functions related to manipulating the inflammatory response.",
keywords = "AnkX, Nucleic acid programmable protein array, PLEKHN1",
author = "Xiaobo Yu and Noll, {Rebecca R.} and {Romero Due{\~n}as}, {Barbara P.} and Allgood, {Samual C.} and Kristi Barker and Caplan, {Jeffrey L.} and Machner, {Matthias P.} and Joshua LaBaer and Ji Qiu and Neunuebel, {M. Ramona}",
year = "2018",
month = "1",
day = "5",
doi = "10.1186/s12866-017-1147-7",
language = "English (US)",
volume = "18",
journal = "BMC Microbiology",
issn = "1471-2180",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Legionella effector AnkX interacts with host nuclear protein PLEKHN1

AU - Yu, Xiaobo

AU - Noll, Rebecca R.

AU - Romero Dueñas, Barbara P.

AU - Allgood, Samual C.

AU - Barker, Kristi

AU - Caplan, Jeffrey L.

AU - Machner, Matthias P.

AU - LaBaer, Joshua

AU - Qiu, Ji

AU - Neunuebel, M. Ramona

PY - 2018/1/5

Y1 - 2018/1/5

N2 - Background: The intracellular bacterial pathogen Legionella pneumophila proliferates in human alveolar macrophages, resulting in a severe pneumonia termed Legionnaires' disease. Throughout the course of infection, L. pneumophila remains enclosed in a specialized membrane compartment that evades fusion with lysosomes. The pathogen delivers over 300 effector proteins into the host cell, altering host pathways in a manner that sets the stage for efficient pathogen replication. The L. pneumophila effector protein AnkX targets host Rab GTPases and functions in preventing fusion of the Legionella-containing vacuole with lysosomes. However, the current understanding of AnkX's interaction with host proteins and the means through which it exerts its cellular function is limited. Results: Here, we investigated the protein interaction network of AnkX by using the nucleic acid programmable protein array (NAPPA), a high-density platform comprising 10,000 unique human ORFs. This approach facilitated the discovery of PLEKHN1 as a novel interaction partner of AnkX. We confirmed this interaction through multiple independent in vitro pull-down, co-immunoprecipitation, and cell-based assays. Structured illumination microscopy revealed that endogenous PLEKHN1 is found in the nucleus and on vesicular compartments, whereas ectopically produced AnkX co-localized with lipid rafts at the plasma membrane. In mammalian cells, HaloTag-AnkX co-localized with endogenous PLEKHN1 on vesicular compartments. A central fragment of AnkX (amino acids 491-809), containing eight ankyrin repeats, extensively co-localized with endogenous PLEKHN1, indicating that this region may harbor a new function. Further, we found that PLEKHN1 associated with multiple proteins involved in the inflammatory response. Conclusions: Altogether, our study provides evidence that in addition to Rab GTPases, the L. pneumophila effector AnkX targets nuclear host proteins and suggests that AnkX may have novel functions related to manipulating the inflammatory response.

AB - Background: The intracellular bacterial pathogen Legionella pneumophila proliferates in human alveolar macrophages, resulting in a severe pneumonia termed Legionnaires' disease. Throughout the course of infection, L. pneumophila remains enclosed in a specialized membrane compartment that evades fusion with lysosomes. The pathogen delivers over 300 effector proteins into the host cell, altering host pathways in a manner that sets the stage for efficient pathogen replication. The L. pneumophila effector protein AnkX targets host Rab GTPases and functions in preventing fusion of the Legionella-containing vacuole with lysosomes. However, the current understanding of AnkX's interaction with host proteins and the means through which it exerts its cellular function is limited. Results: Here, we investigated the protein interaction network of AnkX by using the nucleic acid programmable protein array (NAPPA), a high-density platform comprising 10,000 unique human ORFs. This approach facilitated the discovery of PLEKHN1 as a novel interaction partner of AnkX. We confirmed this interaction through multiple independent in vitro pull-down, co-immunoprecipitation, and cell-based assays. Structured illumination microscopy revealed that endogenous PLEKHN1 is found in the nucleus and on vesicular compartments, whereas ectopically produced AnkX co-localized with lipid rafts at the plasma membrane. In mammalian cells, HaloTag-AnkX co-localized with endogenous PLEKHN1 on vesicular compartments. A central fragment of AnkX (amino acids 491-809), containing eight ankyrin repeats, extensively co-localized with endogenous PLEKHN1, indicating that this region may harbor a new function. Further, we found that PLEKHN1 associated with multiple proteins involved in the inflammatory response. Conclusions: Altogether, our study provides evidence that in addition to Rab GTPases, the L. pneumophila effector AnkX targets nuclear host proteins and suggests that AnkX may have novel functions related to manipulating the inflammatory response.

KW - AnkX

KW - Nucleic acid programmable protein array

KW - PLEKHN1

UR - http://www.scopus.com/inward/record.url?scp=85041963953&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85041963953&partnerID=8YFLogxK

U2 - 10.1186/s12866-017-1147-7

DO - 10.1186/s12866-017-1147-7

M3 - Article

VL - 18

JO - BMC Microbiology

JF - BMC Microbiology

SN - 1471-2180

IS - 1

M1 - 5

ER -