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bioRxiv · 10.1101/2022.09.07.506920

Characterization of TelD, an LXG effector of Streptococcus gallolyticus, antagonized by a non-canonical immunity protein

Teh, W. K.·Ding, Y.·Gubellini, F.·Filloux, A.·Poyart, C.·Dramsi, S.·Givskov, M.
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Abstract

Streptococcus gallolyticus subsp. gallolyticus (SGG) is an opportunistic bacterial pathogen strongly associated with colorectal cancer. Here, through comparative genomics analysis, we demonstrated that the genetic locus encoding the Type VIIb Secretion System (T7SSb) machinery is uniquely present in SGG in two different arrangements. SGG UCN34 carrying the most prevalent T7SSb genetic arrangement was chosen as the reference strain. To identify the effectors secreted by this secretion system, we inactivated the essC gene encoding the motor of this machinery. Comparison of the proteins secreted by UCN34 WT and its isogenic {Delta}essC mutant revealed six T7SSb effector proteins, including the expected WXG effector EsxA and three LXG-containing proteins. In this work, we characterized an LXG-family toxin named herein TelE displaying pore-forming activity. Seven homologs of TelE harboring a conserved glycine zipper motif at the C-terminus were identified in different SGG isolates. Scanning mutagenesis of this motif showed that the glycine residue at position 470 was crucial for TelE pore-forming activity. Unlike other pore-forming toxins commonly antagonized by a membrane protein, TelE activity was antagonized by a small protein TipE belonging to the DUF5085 family. Overall, we report herein a unique SGG T7SSb effector exhibiting a pore-forming activity against non-immune bacteria. IMPORTANCEIn this study, 38 clinical isolates of Streptococcus gallolyticus subsp. gallolyticus (SGG) were sequenced and a genetic locus encoding the Type VIIb secretion system (T7SSb) was found conserved and absent from 16 genomes of the closely related S. gallolyticus subsp. pasteurianus (SGP). The T7SSb is a bona fide pathogenicity island. Here, we report that the model organism SGG strain UCN34 secretes six T7SSb effectors. One of the six effectors named TelE displayed a strong toxicity when overexpressed in Escherichia coli. Our results indicate that TelE is a pore forming toxin whose activity can be antagonized by a non-canonical immunity protein named TipE. Overall, we report a unique toxin-immunity protein pair and our data expand the range of effectors secreted through T7SSb.

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Teh, W. K., Ding, Y., Gubellini, F., Filloux, A., Poyart, C., Dramsi, S., Givskov, M.. 2022-09-07. Characterization of TelD, an LXG effector of Streptococcus gallolyticus, antagonized by a non-canonical immunity protein. https://doi.org/10.1101/2022.09.07.506920

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