Proteoglycan-Dependent Endo-Lysosomal Fusion Affects Intracellular Survival of Salmonella Typhimurium in Epithelial Cells
Proteoglycans (PGs) are glycoconjugates which are predominately expressed on cell surfaces and consist of glycosaminoglycans (GAGs) linked to a core protein. An initial step of GAGs assembly is governed by the β-D-xylosyltransferase enzymes encoded in mammals by the XylT1/XylT2 genes. PGs are essent...
Main Authors: | , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Frontiers Media S.A.
2020-04-01
|
Series: | Frontiers in Immunology |
Subjects: | |
Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2020.00731/full |
id |
doaj-46f37179b8be482696823f3b5a449fe3 |
---|---|
record_format |
Article |
spelling |
doaj-46f37179b8be482696823f3b5a449fe32020-11-25T03:49:29ZengFrontiers Media S.A.Frontiers in Immunology1664-32242020-04-011110.3389/fimmu.2020.00731520125Proteoglycan-Dependent Endo-Lysosomal Fusion Affects Intracellular Survival of Salmonella Typhimurium in Epithelial CellsAlibek Galeev0Abdulhadi Suwandi1Hans Bakker2Ade Oktiviyari3Françoise H. Routier4Lena Krone5Michael Hensel6Guntram A. Grassl7Institute of Medical Microbiology and Hospital Epidemiology, Hannover Medical School and German Center for Infection Research (DZIF), Hanover, GermanyInstitute of Medical Microbiology and Hospital Epidemiology, Hannover Medical School and German Center for Infection Research (DZIF), Hanover, GermanyInstitute of Clinical Biochemistry, Hannover Medical School, Hanover, GermanyInstitute of Medical Microbiology and Hospital Epidemiology, Hannover Medical School and German Center for Infection Research (DZIF), Hanover, GermanyInstitute of Clinical Biochemistry, Hannover Medical School, Hanover, GermanyDivision of Microbiology, University of Osnabrück, Osnabrück, GermanyDivision of Microbiology, University of Osnabrück, Osnabrück, GermanyInstitute of Medical Microbiology and Hospital Epidemiology, Hannover Medical School and German Center for Infection Research (DZIF), Hanover, GermanyProteoglycans (PGs) are glycoconjugates which are predominately expressed on cell surfaces and consist of glycosaminoglycans (GAGs) linked to a core protein. An initial step of GAGs assembly is governed by the β-D-xylosyltransferase enzymes encoded in mammals by the XylT1/XylT2 genes. PGs are essential for the interaction of a cell with other cells as well as with the extracellular matrix. A number of studies highlighted a role of PGs in bacterial adhesion, invasion, and immune response. In this work, we investigated a role of PGs in Salmonella enterica serovar Typhimurium (S. Typhimurium) infection of epithelial cells. Gentamicin protection and chloroquine resistance assays were applied to assess invasion and replication of S. Typhimurium in wild-type and xylosyltransferase-deficient (ΔXylT2) Chinese hamster ovary (CHO) cells lacking PGs. We found that S. Typhimurium adheres to and invades CHO WT and CHO ΔXylT2 cells at comparable levels. However, 24 h after infection, proteoglycan-deficient CHO ΔXylT2 cells are significantly less colonized by S. Typhimurium compared to CHO WT cells. This proteoglycan-dependent phenotype could be rescued by addition of PGs to the cell culture medium, as well as by complementation of the XylT2 gene. Chloroquine resistance assay and immunostaining revealed that in the absence of PGs, significantly less bacteria are associated with Salmonella-containing vacuoles (SCVs) due to a re-distribution of endocytosed gentamicin. Inhibition of endo-lysosomal fusion by a specific inhibitor of phosphatidylinositol phosphate kinase PIKfyve significantly increased S. Typhimurium burden in CHO ΔXylT2 cells demonstrating an important role of PGs for PIKfyve dependent vesicle fusion which is modulated by Salmonella to establish infection. Overall, our results demonstrate that PGs influence survival of intracellular Salmonella in epithelial cells via modulation of PIKfyve-dependent endo-lysosomal fusion.https://www.frontiersin.org/article/10.3389/fimmu.2020.00731/fullSalmonellaproteoglycansglycosaminoglycansxylosyltransferasePIKfyvegentamicin |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Alibek Galeev Abdulhadi Suwandi Hans Bakker Ade Oktiviyari Françoise H. Routier Lena Krone Michael Hensel Guntram A. Grassl |
spellingShingle |
Alibek Galeev Abdulhadi Suwandi Hans Bakker Ade Oktiviyari Françoise H. Routier Lena Krone Michael Hensel Guntram A. Grassl Proteoglycan-Dependent Endo-Lysosomal Fusion Affects Intracellular Survival of Salmonella Typhimurium in Epithelial Cells Frontiers in Immunology Salmonella proteoglycans glycosaminoglycans xylosyltransferase PIKfyve gentamicin |
author_facet |
Alibek Galeev Abdulhadi Suwandi Hans Bakker Ade Oktiviyari Françoise H. Routier Lena Krone Michael Hensel Guntram A. Grassl |
author_sort |
Alibek Galeev |
title |
Proteoglycan-Dependent Endo-Lysosomal Fusion Affects Intracellular Survival of Salmonella Typhimurium in Epithelial Cells |
title_short |
Proteoglycan-Dependent Endo-Lysosomal Fusion Affects Intracellular Survival of Salmonella Typhimurium in Epithelial Cells |
title_full |
Proteoglycan-Dependent Endo-Lysosomal Fusion Affects Intracellular Survival of Salmonella Typhimurium in Epithelial Cells |
title_fullStr |
Proteoglycan-Dependent Endo-Lysosomal Fusion Affects Intracellular Survival of Salmonella Typhimurium in Epithelial Cells |
title_full_unstemmed |
Proteoglycan-Dependent Endo-Lysosomal Fusion Affects Intracellular Survival of Salmonella Typhimurium in Epithelial Cells |
title_sort |
proteoglycan-dependent endo-lysosomal fusion affects intracellular survival of salmonella typhimurium in epithelial cells |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2020-04-01 |
description |
Proteoglycans (PGs) are glycoconjugates which are predominately expressed on cell surfaces and consist of glycosaminoglycans (GAGs) linked to a core protein. An initial step of GAGs assembly is governed by the β-D-xylosyltransferase enzymes encoded in mammals by the XylT1/XylT2 genes. PGs are essential for the interaction of a cell with other cells as well as with the extracellular matrix. A number of studies highlighted a role of PGs in bacterial adhesion, invasion, and immune response. In this work, we investigated a role of PGs in Salmonella enterica serovar Typhimurium (S. Typhimurium) infection of epithelial cells. Gentamicin protection and chloroquine resistance assays were applied to assess invasion and replication of S. Typhimurium in wild-type and xylosyltransferase-deficient (ΔXylT2) Chinese hamster ovary (CHO) cells lacking PGs. We found that S. Typhimurium adheres to and invades CHO WT and CHO ΔXylT2 cells at comparable levels. However, 24 h after infection, proteoglycan-deficient CHO ΔXylT2 cells are significantly less colonized by S. Typhimurium compared to CHO WT cells. This proteoglycan-dependent phenotype could be rescued by addition of PGs to the cell culture medium, as well as by complementation of the XylT2 gene. Chloroquine resistance assay and immunostaining revealed that in the absence of PGs, significantly less bacteria are associated with Salmonella-containing vacuoles (SCVs) due to a re-distribution of endocytosed gentamicin. Inhibition of endo-lysosomal fusion by a specific inhibitor of phosphatidylinositol phosphate kinase PIKfyve significantly increased S. Typhimurium burden in CHO ΔXylT2 cells demonstrating an important role of PGs for PIKfyve dependent vesicle fusion which is modulated by Salmonella to establish infection. Overall, our results demonstrate that PGs influence survival of intracellular Salmonella in epithelial cells via modulation of PIKfyve-dependent endo-lysosomal fusion. |
topic |
Salmonella proteoglycans glycosaminoglycans xylosyltransferase PIKfyve gentamicin |
url |
https://www.frontiersin.org/article/10.3389/fimmu.2020.00731/full |
work_keys_str_mv |
AT alibekgaleev proteoglycandependentendolysosomalfusionaffectsintracellularsurvivalofsalmonellatyphimuriuminepithelialcells AT abdulhadisuwandi proteoglycandependentendolysosomalfusionaffectsintracellularsurvivalofsalmonellatyphimuriuminepithelialcells AT hansbakker proteoglycandependentendolysosomalfusionaffectsintracellularsurvivalofsalmonellatyphimuriuminepithelialcells AT adeoktiviyari proteoglycandependentendolysosomalfusionaffectsintracellularsurvivalofsalmonellatyphimuriuminepithelialcells AT francoisehroutier proteoglycandependentendolysosomalfusionaffectsintracellularsurvivalofsalmonellatyphimuriuminepithelialcells AT lenakrone proteoglycandependentendolysosomalfusionaffectsintracellularsurvivalofsalmonellatyphimuriuminepithelialcells AT michaelhensel proteoglycandependentendolysosomalfusionaffectsintracellularsurvivalofsalmonellatyphimuriuminepithelialcells AT guntramagrassl proteoglycandependentendolysosomalfusionaffectsintracellularsurvivalofsalmonellatyphimuriuminepithelialcells |
_version_ |
1724495162022821888 |