Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies
Gram-negative pathogens evolved a syringe-like nanomachine, termed type 3 secretion system, to deliver protein effectors into the cytoplasm of host cells. An essential component of this system is a long helical needle filament that protrudes from the bacterial surface and connects the cytoplasms of...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-09-01
|
| Series: | Biochemistry and Biophysics Reports |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580821001333 |
| _version_ | 1819123370830069760 |
|---|---|
| author | Vadim Kotov Michele Lunelli Jiri Wald Michael Kolbe Thomas C. Marlovits |
| author_facet | Vadim Kotov Michele Lunelli Jiri Wald Michael Kolbe Thomas C. Marlovits |
| author_sort | Vadim Kotov |
| collection | DOAJ |
| description | Gram-negative pathogens evolved a syringe-like nanomachine, termed type 3 secretion system, to deliver protein effectors into the cytoplasm of host cells. An essential component of this system is a long helical needle filament that protrudes from the bacterial surface and connects the cytoplasms of the bacterium and the eukaryotic cell. Previous structural research was predominantly focused on reconstituted type 3 needle filaments, which lacked the biological context. In this work we introduce a facile procedure to obtain high-resolution cryo-EM structure of needle filaments attached to the basal body of type 3 secretion systems. We validate our approach by solving the structure of Salmonella PrgI filament and demonstrate its utility by obtaining the first high-resolution cryo-EM reconstruction of Shigella MxiH filament. Our work paves the way to systematic structural characterization of attached type 3 needle filaments in the context of mutagenesis studies, protein structural evolution and drug development. |
| first_indexed | 2024-12-22T07:07:17Z |
| format | Article |
| id | doaj.art-a8f59de1bfad421ea6a2c5db03cc3681 |
| institution | Directory Open Access Journal |
| issn | 2405-5808 |
| language | English |
| last_indexed | 2024-12-22T07:07:17Z |
| publishDate | 2021-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj.art-a8f59de1bfad421ea6a2c5db03cc36812022-12-21T18:34:38ZengElsevierBiochemistry and Biophysics Reports2405-58082021-09-0127101039Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodiesVadim Kotov0Michele Lunelli1Jiri Wald2Michael Kolbe3Thomas C. Marlovits4Centre for Structural Systems Biology (CSSB), Notkestraße 85, 22607 Hamburg, Germany; University Medical Centre Hamburg-Eppendorf (UKE), Martinistraße 52, 20246 Hamburg, Germany; Deutsches Elektronen-Synchrotron (DESY), Notkestraße 85, 22607 Hamburg, GermanyCentre for Structural Systems Biology (CSSB), Notkestraße 85, 22607 Hamburg, Germany; Department of Structural Infection Biology, Helmholtz Centre for Infection Research (HZI), Inhoffenstraße 7, 38124 Braunschweig, GermanyCentre for Structural Systems Biology (CSSB), Notkestraße 85, 22607 Hamburg, Germany; University Medical Centre Hamburg-Eppendorf (UKE), Martinistraße 52, 20246 Hamburg, Germany; Deutsches Elektronen-Synchrotron (DESY), Notkestraße 85, 22607 Hamburg, GermanyCentre for Structural Systems Biology (CSSB), Notkestraße 85, 22607 Hamburg, Germany; Department of Structural Infection Biology, Helmholtz Centre for Infection Research (HZI), Inhoffenstraße 7, 38124 Braunschweig, Germany; Department of Chemistry, Faculty of Mathematics, Informatics and Natural Sciences, University of Hamburg, Rothenbaumchaussee 19, 20148 Hamburg, Germany; Corresponding author. Centre for Structural Systems Biology (CSSB), Notkestraße 85, 22607 Hamburg, Germany.Centre for Structural Systems Biology (CSSB), Notkestraße 85, 22607 Hamburg, Germany; University Medical Centre Hamburg-Eppendorf (UKE), Martinistraße 52, 20246 Hamburg, Germany; Deutsches Elektronen-Synchrotron (DESY), Notkestraße 85, 22607 Hamburg, Germany; Corresponding author. Centre for Structural Systems Biology (CSSB), Notkestraße 85, 22607 Hamburg, Germany.Gram-negative pathogens evolved a syringe-like nanomachine, termed type 3 secretion system, to deliver protein effectors into the cytoplasm of host cells. An essential component of this system is a long helical needle filament that protrudes from the bacterial surface and connects the cytoplasms of the bacterium and the eukaryotic cell. Previous structural research was predominantly focused on reconstituted type 3 needle filaments, which lacked the biological context. In this work we introduce a facile procedure to obtain high-resolution cryo-EM structure of needle filaments attached to the basal body of type 3 secretion systems. We validate our approach by solving the structure of Salmonella PrgI filament and demonstrate its utility by obtaining the first high-resolution cryo-EM reconstruction of Shigella MxiH filament. Our work paves the way to systematic structural characterization of attached type 3 needle filaments in the context of mutagenesis studies, protein structural evolution and drug development.http://www.sciencedirect.com/science/article/pii/S2405580821001333Type 3 secretion systemCryo electron microscopyNeedle filamentHelical reconstructionShigellaSalmonella, host-pathogen interaction |
| spellingShingle | Vadim Kotov Michele Lunelli Jiri Wald Michael Kolbe Thomas C. Marlovits Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies Biochemistry and Biophysics Reports Type 3 secretion system Cryo electron microscopy Needle filament Helical reconstruction Shigella Salmonella, host-pathogen interaction |
| title | Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies |
| title_full | Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies |
| title_fullStr | Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies |
| title_full_unstemmed | Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies |
| title_short | Helical reconstruction of Salmonella and Shigella needle filaments attached to type 3 basal bodies |
| title_sort | helical reconstruction of salmonella and shigella needle filaments attached to type 3 basal bodies |
| topic | Type 3 secretion system Cryo electron microscopy Needle filament Helical reconstruction Shigella Salmonella, host-pathogen interaction |
| url | http://www.sciencedirect.com/science/article/pii/S2405580821001333 |
| work_keys_str_mv | AT vadimkotov helicalreconstructionofsalmonellaandshigellaneedlefilamentsattachedtotype3basalbodies AT michelelunelli helicalreconstructionofsalmonellaandshigellaneedlefilamentsattachedtotype3basalbodies AT jiriwald helicalreconstructionofsalmonellaandshigellaneedlefilamentsattachedtotype3basalbodies AT michaelkolbe helicalreconstructionofsalmonellaandshigellaneedlefilamentsattachedtotype3basalbodies AT thomascmarlovits helicalreconstructionofsalmonellaandshigellaneedlefilamentsattachedtotype3basalbodies |