The breadth and molecular basis of hcp-driven type vi secretion system effector delivery

Sophie A. Howard, R. Christopher D. Furniss, Dora Bonini, Himani Amin, Patricia Paracuellos, David Zlotkin, Tiago R.D. Costa, Asaf Levy, Despoina A.I. Mavridou, Alain Filloux*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


The type VI secretion system (T6SS) is a bacterial nanoscale weapon that delivers toxins into prey ranging from bacteria and fungi to animal hosts. The cytosolic contractile sheath of the system wraps around stacked hexameric rings of Hcp proteins, which form an inner tube. At the tip of this tube is a puncturing device comprising a trimeric VgrG topped by a monomeric PAAR protein. The number of toxins a single system delivers per firing event remains unknown, since effectors can be loaded on diverse sites of the T6SS apparatus, notably the inner tube and the puncturing device. Each VgrG or PAAR can bind one effector, and additional effector cargoes can be carried in the Hcp ring lumen. While many VgrG-and PAAR-bound toxins have been characterized, to date, very few Hcp-bound effectors are known. Here, we used 3 known Pseudomonas aeruginosa Hcp proteins (Hcp1 to-3), each of which associates with one of the three T6SSs in this organism (H1-T6SS, H2-T6SS, and H3-T6SS), to perform in vivo pulldown assays. We confirmed the known interactions of Hcp1 with Tse1 to-4, further copurified a Hcp1-Tse4 complex, and identified potential novel Hcp1-bound effectors. Moreover, we demonstrated that Hcp2 and Hcp3 can shuttle T6SS cargoes toxic to Escherichia coli. Finally, we used a Tse1-Bla chimera to probe the loading strategy for Hcp passengers and found that while large effectors can be loaded onto Hcp, the formed complex jams the system, abrogating T6SS function.

Original languageAmerican English
Article numbere00262-21
Issue number3
StatePublished - 2021
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2021 Howard et al.


  • Hcp
  • Protein secretion
  • Pseudomonas
  • T6SS
  • Toxin


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