Interactions of the Transmembrane Polymeric Rings of the <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Type III Secretion System
ABSTRACT The type III secretion system (T3SS) is an interspecies protein transport machine that plays a major role in interactions of Gram-negative bacteria with animals and plants by delivering bacterial effector proteins into host cells. T3SSs span both membranes of Gram-negative bacteria by formi...
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American Society for Microbiology
2010
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oai:doaj.org-article:c5b0c9a13dc04987adec5af22be53b0f2021-11-15T15:38:16ZInteractions of the Transmembrane Polymeric Rings of the <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Type III Secretion System10.1128/mBio.00158-102150-7511https://doaj.org/article/c5b0c9a13dc04987adec5af22be53b0f2010-08-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00158-10https://doaj.org/toc/2150-7511ABSTRACT The type III secretion system (T3SS) is an interspecies protein transport machine that plays a major role in interactions of Gram-negative bacteria with animals and plants by delivering bacterial effector proteins into host cells. T3SSs span both membranes of Gram-negative bacteria by forming a structure of connected oligomeric rings termed the needle complex (NC). Here, the localization of subunits in the Salmonella enterica serovar Typhimurium T3SS NC were probed via mass spectrometry-assisted identification of chemical cross-links in intact NC preparations. Cross-links between amino acids near the amino terminus of the outer membrane ring component InvG and the carboxyl terminus of the inner membrane ring component PrgH and between the two inner membrane components PrgH and PrgK allowed for spatial localization of the three ring components within the electron density map structures of NCs. Mutational and biochemical analysis demonstrated that the amino terminus of InvG and the carboxyl terminus of PrgH play a critical role in the assembly and function of the T3SS apparatus. Analysis of an InvG mutant indicates that the structure of the InvG oligomer can affect the switching of the T3SS substrate to translocon and effector components. This study provides insights into how structural organization of needle complex base components promotes T3SS assembly and function. IMPORTANCE Many biological macromolecular complexes are composed of symmetrical homomers, which assemble into larger structures. Some complexes, such as secretion systems, span biological membranes, forming hydrophilic domains to move substrates across lipid bilayers. Type III secretion systems (T3SSs) deliver bacterial effector proteins directly to the host cell cytoplasm and allow for critical interactions between many Gram-negative pathogenic bacterial species and their hosts. Progress has been made towards the goal of determining the three-dimensional structure of the secretion apparatus by determination of high-resolution crystal structures of individual protein subunits and low-resolution models of the assembly, using reconstructions of cryoelectron microscopy images. However, a more refined picture of the localization of periplasmic ring structures within the assembly and their interactions has only recently begun to emerge. This work localizes T3SS transmembrane rings and identifies structural elements that affect substrate switching and are essential to the assembly of components that are inserted into host cell membranes.Sarah SanowarPragya SinghRichard A. PfuetznerIngemar AndréHongjin ZhengThomas SpreterNatalie C. J. StrynadkaTamir GonenDavid BakerDavid R. GoodlettSamuel I. MillerAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 1, Iss 3 (2010) |
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Microbiology QR1-502 |
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Microbiology QR1-502 Sarah Sanowar Pragya Singh Richard A. Pfuetzner Ingemar André Hongjin Zheng Thomas Spreter Natalie C. J. Strynadka Tamir Gonen David Baker David R. Goodlett Samuel I. Miller Interactions of the Transmembrane Polymeric Rings of the <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Type III Secretion System |
| description |
ABSTRACT The type III secretion system (T3SS) is an interspecies protein transport machine that plays a major role in interactions of Gram-negative bacteria with animals and plants by delivering bacterial effector proteins into host cells. T3SSs span both membranes of Gram-negative bacteria by forming a structure of connected oligomeric rings termed the needle complex (NC). Here, the localization of subunits in the Salmonella enterica serovar Typhimurium T3SS NC were probed via mass spectrometry-assisted identification of chemical cross-links in intact NC preparations. Cross-links between amino acids near the amino terminus of the outer membrane ring component InvG and the carboxyl terminus of the inner membrane ring component PrgH and between the two inner membrane components PrgH and PrgK allowed for spatial localization of the three ring components within the electron density map structures of NCs. Mutational and biochemical analysis demonstrated that the amino terminus of InvG and the carboxyl terminus of PrgH play a critical role in the assembly and function of the T3SS apparatus. Analysis of an InvG mutant indicates that the structure of the InvG oligomer can affect the switching of the T3SS substrate to translocon and effector components. This study provides insights into how structural organization of needle complex base components promotes T3SS assembly and function. IMPORTANCE Many biological macromolecular complexes are composed of symmetrical homomers, which assemble into larger structures. Some complexes, such as secretion systems, span biological membranes, forming hydrophilic domains to move substrates across lipid bilayers. Type III secretion systems (T3SSs) deliver bacterial effector proteins directly to the host cell cytoplasm and allow for critical interactions between many Gram-negative pathogenic bacterial species and their hosts. Progress has been made towards the goal of determining the three-dimensional structure of the secretion apparatus by determination of high-resolution crystal structures of individual protein subunits and low-resolution models of the assembly, using reconstructions of cryoelectron microscopy images. However, a more refined picture of the localization of periplasmic ring structures within the assembly and their interactions has only recently begun to emerge. This work localizes T3SS transmembrane rings and identifies structural elements that affect substrate switching and are essential to the assembly of components that are inserted into host cell membranes. |
| format |
article |
| author |
Sarah Sanowar Pragya Singh Richard A. Pfuetzner Ingemar André Hongjin Zheng Thomas Spreter Natalie C. J. Strynadka Tamir Gonen David Baker David R. Goodlett Samuel I. Miller |
| author_facet |
Sarah Sanowar Pragya Singh Richard A. Pfuetzner Ingemar André Hongjin Zheng Thomas Spreter Natalie C. J. Strynadka Tamir Gonen David Baker David R. Goodlett Samuel I. Miller |
| author_sort |
Sarah Sanowar |
| title |
Interactions of the Transmembrane Polymeric Rings of the <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Type III Secretion System |
| title_short |
Interactions of the Transmembrane Polymeric Rings of the <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Type III Secretion System |
| title_full |
Interactions of the Transmembrane Polymeric Rings of the <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Type III Secretion System |
| title_fullStr |
Interactions of the Transmembrane Polymeric Rings of the <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Type III Secretion System |
| title_full_unstemmed |
Interactions of the Transmembrane Polymeric Rings of the <named-content content-type="genus-species">Salmonella enterica</named-content> Serovar Typhimurium Type III Secretion System |
| title_sort |
interactions of the transmembrane polymeric rings of the <named-content content-type="genus-species">salmonella enterica</named-content> serovar typhimurium type iii secretion system |
| publisher |
American Society for Microbiology |
| publishDate |
2010 |
| url |
https://doaj.org/article/c5b0c9a13dc04987adec5af22be53b0f |
| work_keys_str_mv |
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1718427852605489152 |