Rhs NADase effectors and their immunity proteins are exchangeable mediators of inter-bacterial competition in Serratia

Martin Hagan, Genady Pankov, Ramses Gallegos-Monterrosa, David J. Williams, Christopher Earl, Grant Buchanan, William N. Hunter () and Sarah J. Coulthurst ()
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Martin Hagan: University of Dundee
Genady Pankov: University of Dundee
Ramses Gallegos-Monterrosa: University of Dundee
David J. Williams: University of Dundee
Christopher Earl: University of Dundee
Grant Buchanan: University of Dundee
William N. Hunter: University of Dundee
Sarah J. Coulthurst: University of Dundee

Nature Communications, 2023, vol. 14, issue 1, 1-16

Abstract: Abstract Many bacterial species use Type VI secretion systems (T6SSs) to deliver anti-bacterial effector proteins into neighbouring bacterial cells, representing an important mechanism of inter-bacterial competition. Specific immunity proteins protect bacteria from the toxic action of their own effectors, whilst orphan immunity proteins without a cognate effector may provide protection against incoming effectors from non-self competitors. T6SS-dependent Rhs effectors contain a variable C-terminal toxin domain (CT), with the cognate immunity protein encoded immediately downstream of the effector. Here, we demonstrate that Rhs1 effectors from two strains of Serratia marcescens, the model strain Db10 and clinical isolate SJC1036, possess distinct CTs which both display NAD(P)+ glycohydrolase activity but belong to different subgroups of NADase from each other and other T6SS-associated NADases. Comparative structural analysis identifies conserved functions required for NADase activity and reveals that unrelated NADase immunity proteins utilise a common mechanism of effector inhibition. By replicating a natural recombination event, we show successful functional exchange of CTs and demonstrate that Db10 encodes an orphan immunity protein which provides protection against T6SS-delivered SJC1036 NADase. Our findings highlight the flexible use of Rhs effectors and orphan immunity proteins during inter-strain competition and the repeated adoption of NADase toxins as weapons against bacterial cells.

Date: 2023
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DOI: 10.1038/s41467-023-41751-3

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