Haemolysin Coregulated Protein Is an Exported Receptor and Chaperone of Type VI Secretion Substrates

•Hcp is a chaperone required for the intracellular accumulation of Tse2•Export of type VI secretion effectors requires direct interaction with the pore of Hcp•Cognate Hcp-effector interactions can define substrate specificity of the T6SS•Substrate recognition and energization of secretion are uncoupled in the T6SS

SummarySecretion systems require high-fidelity mechanisms to discriminate substrates among the vast cytoplasmic pool of proteins. Factors mediating substrate recognition by the type VI secretion system (T6SS) of Gram-negative bacteria, a widespread pathway that translocates effector proteins into target bacterial cells, have not been defined. We report that haemolysin coregulated protein (Hcp), a ring-shaped hexamer secreted by all characterized T6SSs, binds specifically to cognate effector molecules. Electron microscopy analysis of an Hcp-effector complex from Pseudomonas aeruginosa revealed the effector bound to the inner surface of Hcp. Further studies demonstrated that interaction with the Hcp pore is a general requirement for secretion of diverse effectors encompassing several enzymatic classes. Though previous models depict Hcp as a static conduit, our data indicate it is a chaperone and receptor of substrates. These unique functions of a secreted protein highlight fundamental differences between the export mechanism of T6 and other characterized secretory pathways.