TssK is a trimeric cytoplasmic protein interacting with components of both phage-like and membrane anchoring complexes of the type VI secretion system.
The Type VI secretion system (T6SS) is a macromolecular machine that mediates bacteria-host or bacteria-bacteria interactions. The T6SS core apparatus assembles from 13 proteins that form two sub-assemblies: a phage-like complex and a trans-envelope complex. The Hcp, VgrG, TssE, and TssB/C subunits are structurally and functionally related to components of the tail of contractile bacteriophages. This phage-like structure is thought to be anchored to the membrane by a trans-envelope complex composed of the TssJ, TssL, and TssM proteins. However, how the two sub-complexes are connected remains unknown. Here we identify TssK, a protein that establishes contacts with the two T6SS sub-complexes through direct interactions with TssL, Hcp, and TssC. TssK is a cytoplasmic protein assembling trimers that display a three-armed shape, as revealed by TEM and SAXS analyses. Fluorescence microscopy experiments further demonstrate the requirement of TssK for sheath assembly. Our results suggest a central role for TssK by linking both complexes during T6SS assembly.
Laboratoire d'Ingénierie des Systèmes Macromoléculaires, Institut de Microbiologie de la Méditerranée, CNRS, UMR 7255, Aix-Marseille Université, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
Abdelrahim
A
Zoued
Architecture et Fonction des Macromolécules Biologiques, Campus de Luminy, Case 932, Aix-Marseille Université, CNRS UMR 6098, 13288 Marseille Cedex 09, France.
Eric
E
Durand
Architecture et Fonction des Macromolécules Biologiques, Campus de Luminy, Case 932, Aix-Marseille Université, CNRS UMR 6098, 13288 Marseille Cedex 09, France.
Cecilia
C
Bebeacua
Laboratoire d'Ingénierie des Systèmes Macromoléculaires, Institut de Microbiologie de la Méditerranée, CNRS, UMR 7255, Aix-Marseille Université, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
Yannick R
YR
Brunet
Architecture et Fonction des Macromolécules Biologiques, Campus de Luminy, Case 932, Aix-Marseille Université, CNRS UMR 6098, 13288 Marseille Cedex 09, France.
Badreddine
B
Douzi
Architecture et Fonction des Macromolécules Biologiques, Campus de Luminy, Case 932, Aix-Marseille Université, CNRS UMR 6098, 13288 Marseille Cedex 09, France.
Christian
C
Cambillau
Laboratoire d'Ingénierie des Systèmes Macromoléculaires, Institut de Microbiologie de la Méditerranée, CNRS, UMR 7255, Aix-Marseille Université, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France.
Eric
E
Cascales
Laboratoire d'Ingénierie des Systèmes Macromoléculaires, Institut de Microbiologie de la Méditerranée, CNRS, UMR 7255, Aix-Marseille Université, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France. Electronic address: ljournet@imm.cnrs.fr.
Laure
L
Journet
D058947:Bacterial Secretion Systems / Q000502:physiology*; D001435:Bacteriophages; D048168:Escherichia coli K12 / Q000235:genetics / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D046912:Multiprotein Complexes / Q000235:genetics / Q000378:metabolism*
D058947:Bacterial Secretion Systems; D029968:Escherichia coli Proteins; D046912:Multiprotein Complexes
Bacterial Pathogenesis; Hcp; Membrane Transport; Microbiology; Protein Complexes; Protein Secretion; Sheath Assembly; TssK; TssL; Type VI Secretion