Selective Association of Outer Surface Lipoproteins with the Lipid Rafts of <named-content content-type="genus-species">Borrelia burgdorferi</named-content>

ABSTRACT Borrelia burgdorferi contains unique cholesterol-glycolipid-rich lipid rafts that are associated with lipoproteins. These complexes suggest the existence of macromolecular structures that have not been reported for prokaryotes. Outer surface lipoproteins OspA, OspB, and OspC were studied fo...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Alvaro Toledo, Jameson T. Crowley, James L. Coleman, Timothy J. LaRocca, Salvatore Chiantia, Erwin London, Jorge L. Benach
Formato: article
Lenguaje:EN
Publicado: American Society for Microbiology 2014
Materias:
Acceso en línea:https://doaj.org/article/c93f466388814badbe034779d2a066b9
Etiquetas: Agregar Etiqueta
Sin Etiquetas, Sea el primero en etiquetar este registro!
id oai:doaj.org-article:c93f466388814badbe034779d2a066b9
record_format dspace
spelling oai:doaj.org-article:c93f466388814badbe034779d2a066b92021-11-15T15:45:13ZSelective Association of Outer Surface Lipoproteins with the Lipid Rafts of <named-content content-type="genus-species">Borrelia burgdorferi</named-content>10.1128/mBio.00899-142150-7511https://doaj.org/article/c93f466388814badbe034779d2a066b92014-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00899-14https://doaj.org/toc/2150-7511ABSTRACT Borrelia burgdorferi contains unique cholesterol-glycolipid-rich lipid rafts that are associated with lipoproteins. These complexes suggest the existence of macromolecular structures that have not been reported for prokaryotes. Outer surface lipoproteins OspA, OspB, and OspC were studied for their participation in the formation of lipid rafts. Single-gene deletion mutants with deletions of ∆ospA, ∆ospB, and ∆ospC and a spontaneous gene mutant, strain B313, which does not express OspA and OspB, were used to establish their structural roles in the lipid rafts. All mutant strains used in this study produced detergent-resistant membranes, a common characteristic of lipid rafts, and had similar lipid and protein slot blot profiles. Lipoproteins OspA and OspB but not OspC were shown to be associated with lipid rafts by transmission electron microscopy. When the ability to form lipid rafts in live B. burgdorferi spirochetes was measured by fluorescence resonance energy transfer (FRET), strain B313 showed a statistically significant lower level of segregation into ordered and disordered membrane domains than did the wild-type and the other single-deletion mutants. The transformation of a B313 strain with a shuttle plasmid containing ospA restored the phenotype shared by the wild type and the single-deletion mutants, demonstrating that OspA and OspB have redundant functions. In contrast, a transformed B313 overexpressing OspC neither rescued the FRET nor colocalized with the lipid rafts. Because these lipoproteins are expressed at different stages of the life cycle of B. burgdorferi, their selective association is likely to have an important role in the structure of prokaryotic lipid rafts and in the organism’s adaptation to changing environments. IMPORTANCE Lipid rafts are cholesterol-rich clusters within the membranes of cells. Lipid rafts contain proteins that have functions in sensing the cell environment and transmitting signals. Although selective proteins are present in lipid rafts, little is known about their structural contribution to these domains. Borrelia burgdorferi, the agent of Lyme disease, has lipid rafts, which are novel structures in bacteria. Here, we have shown that the raft-associated lipoproteins OspA and OspB selectively contribute to lipid rafts. A similar but non-raft-associated lipoprotein, OspC, cannot substitute for the role of OspA and OspB. In this study, we have demonstrated that lipoprotein association with lipid rafts is selective, further suggesting a functional adaptation to different stages of the spirochete life cycle. The results of this study are of broader importance and can serve as a model for other bacteria that also possess cholesterol in their membranes and, therefore, may share lipid raft traits with Borrelia.Alvaro ToledoJameson T. CrowleyJames L. ColemanTimothy J. LaRoccaSalvatore ChiantiaErwin LondonJorge L. BenachAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 5, Iss 2 (2014)
institution DOAJ
collection DOAJ
language EN
topic Microbiology
QR1-502
spellingShingle Microbiology
QR1-502
Alvaro Toledo
Jameson T. Crowley
James L. Coleman
Timothy J. LaRocca
Salvatore Chiantia
Erwin London
Jorge L. Benach
Selective Association of Outer Surface Lipoproteins with the Lipid Rafts of <named-content content-type="genus-species">Borrelia burgdorferi</named-content>
description ABSTRACT Borrelia burgdorferi contains unique cholesterol-glycolipid-rich lipid rafts that are associated with lipoproteins. These complexes suggest the existence of macromolecular structures that have not been reported for prokaryotes. Outer surface lipoproteins OspA, OspB, and OspC were studied for their participation in the formation of lipid rafts. Single-gene deletion mutants with deletions of ∆ospA, ∆ospB, and ∆ospC and a spontaneous gene mutant, strain B313, which does not express OspA and OspB, were used to establish their structural roles in the lipid rafts. All mutant strains used in this study produced detergent-resistant membranes, a common characteristic of lipid rafts, and had similar lipid and protein slot blot profiles. Lipoproteins OspA and OspB but not OspC were shown to be associated with lipid rafts by transmission electron microscopy. When the ability to form lipid rafts in live B. burgdorferi spirochetes was measured by fluorescence resonance energy transfer (FRET), strain B313 showed a statistically significant lower level of segregation into ordered and disordered membrane domains than did the wild-type and the other single-deletion mutants. The transformation of a B313 strain with a shuttle plasmid containing ospA restored the phenotype shared by the wild type and the single-deletion mutants, demonstrating that OspA and OspB have redundant functions. In contrast, a transformed B313 overexpressing OspC neither rescued the FRET nor colocalized with the lipid rafts. Because these lipoproteins are expressed at different stages of the life cycle of B. burgdorferi, their selective association is likely to have an important role in the structure of prokaryotic lipid rafts and in the organism’s adaptation to changing environments. IMPORTANCE Lipid rafts are cholesterol-rich clusters within the membranes of cells. Lipid rafts contain proteins that have functions in sensing the cell environment and transmitting signals. Although selective proteins are present in lipid rafts, little is known about their structural contribution to these domains. Borrelia burgdorferi, the agent of Lyme disease, has lipid rafts, which are novel structures in bacteria. Here, we have shown that the raft-associated lipoproteins OspA and OspB selectively contribute to lipid rafts. A similar but non-raft-associated lipoprotein, OspC, cannot substitute for the role of OspA and OspB. In this study, we have demonstrated that lipoprotein association with lipid rafts is selective, further suggesting a functional adaptation to different stages of the spirochete life cycle. The results of this study are of broader importance and can serve as a model for other bacteria that also possess cholesterol in their membranes and, therefore, may share lipid raft traits with Borrelia.
format article
author Alvaro Toledo
Jameson T. Crowley
James L. Coleman
Timothy J. LaRocca
Salvatore Chiantia
Erwin London
Jorge L. Benach
author_facet Alvaro Toledo
Jameson T. Crowley
James L. Coleman
Timothy J. LaRocca
Salvatore Chiantia
Erwin London
Jorge L. Benach
author_sort Alvaro Toledo
title Selective Association of Outer Surface Lipoproteins with the Lipid Rafts of <named-content content-type="genus-species">Borrelia burgdorferi</named-content>
title_short Selective Association of Outer Surface Lipoproteins with the Lipid Rafts of <named-content content-type="genus-species">Borrelia burgdorferi</named-content>
title_full Selective Association of Outer Surface Lipoproteins with the Lipid Rafts of <named-content content-type="genus-species">Borrelia burgdorferi</named-content>
title_fullStr Selective Association of Outer Surface Lipoproteins with the Lipid Rafts of <named-content content-type="genus-species">Borrelia burgdorferi</named-content>
title_full_unstemmed Selective Association of Outer Surface Lipoproteins with the Lipid Rafts of <named-content content-type="genus-species">Borrelia burgdorferi</named-content>
title_sort selective association of outer surface lipoproteins with the lipid rafts of <named-content content-type="genus-species">borrelia burgdorferi</named-content>
publisher American Society for Microbiology
publishDate 2014
url https://doaj.org/article/c93f466388814badbe034779d2a066b9
work_keys_str_mv AT alvarotoledo selectiveassociationofoutersurfacelipoproteinswiththelipidraftsofnamedcontentcontenttypegenusspeciesborreliaburgdorferinamedcontent
AT jamesontcrowley selectiveassociationofoutersurfacelipoproteinswiththelipidraftsofnamedcontentcontenttypegenusspeciesborreliaburgdorferinamedcontent
AT jameslcoleman selectiveassociationofoutersurfacelipoproteinswiththelipidraftsofnamedcontentcontenttypegenusspeciesborreliaburgdorferinamedcontent
AT timothyjlarocca selectiveassociationofoutersurfacelipoproteinswiththelipidraftsofnamedcontentcontenttypegenusspeciesborreliaburgdorferinamedcontent
AT salvatorechiantia selectiveassociationofoutersurfacelipoproteinswiththelipidraftsofnamedcontentcontenttypegenusspeciesborreliaburgdorferinamedcontent
AT erwinlondon selectiveassociationofoutersurfacelipoproteinswiththelipidraftsofnamedcontentcontenttypegenusspeciesborreliaburgdorferinamedcontent
AT jorgelbenach selectiveassociationofoutersurfacelipoproteinswiththelipidraftsofnamedcontentcontenttypegenusspeciesborreliaburgdorferinamedcontent
_version_ 1718427611760164864