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Identification of a target cell permissive factor required for contact-dependent growth inhibition (CDI)

Identification of a target cell permissive factor required for contact-dependent growth inhibition (CDI)
Identification of a target cell permissive factor required for contact-dependent growth inhibition (CDI)
Bacterial contact-dependent growth inhibition (CDI) is mediated by the CdiB/CdiA family of two-partner secretion proteins. CdiA effector proteins are exported onto the surface of CDI(+) inhibitor cells, where they interact with susceptible bacteria and deliver effectors/toxins derived from their C-terminal regions (CdiA-CT). CDI(+) cells also produce an immunity protein that binds the CdiA-CT and blocks its activity to prevent autoinhibition. Here, we show that the CdiA-CT from uropathogenic Escherichia coli strain 536 (UPEC536) is a latent tRNase that requires activation by the biosynthetic enzyme CysK (O-acetylserine sulfhydrylase A). UPEC536 CdiA-CT exhibits no nuclease activity in vitro, but cleaves within transfer RNA (tRNA) anti-codon loops when purified CysK is added. CysK and CdiA-CT form a stable complex, and their binding interaction appears to mimic that of the CysK/CysE cysteine synthase complex. CdiA-CT activation is also required for growth inhibition. Synthesis of CdiA-CT in E. coli cysK(+) cells arrests cell growth, whereas the growth of ΔcysK mutants is unaffected by the toxin. Moreover, E. coli ΔcysK cells are completely resistant to inhibitor cells expressing UPEC536 CdiA, indicating that CysK is required to activate the tRNase during CDI. Thus, CysK acts as a permissive factor for CDI, providing a potential mechanism to modulate growth inhibition in target cells.
0890-9369
515-525
Webb, Jeremy
ec0a5c4e-86cc-4ae9-b390-7298f5d65f8d
Diner, Elie
86338b19-73b1-41bb-b393-41f6f5e2ef32
Beck, Christina
b31fde2c-9e69-4ab9-802c-e09166e36251
Low, David A.
ba5ab1db-5817-4871-8905-47e5ba8de26c
Hayes, Christopher S.
d4d6cee7-197b-46c8-a822-e120f81b24e6
Webb, Jeremy
ec0a5c4e-86cc-4ae9-b390-7298f5d65f8d
Diner, Elie
86338b19-73b1-41bb-b393-41f6f5e2ef32
Beck, Christina
b31fde2c-9e69-4ab9-802c-e09166e36251
Low, David A.
ba5ab1db-5817-4871-8905-47e5ba8de26c
Hayes, Christopher S.
d4d6cee7-197b-46c8-a822-e120f81b24e6

Webb, Jeremy, Diner, Elie, Beck, Christina, Low, David A. and Hayes, Christopher S. (2012) Identification of a target cell permissive factor required for contact-dependent growth inhibition (CDI). Genes & Development, 26, 515-525. (doi:10.1101/gad.182345.111).

Record type: Article

Abstract

Bacterial contact-dependent growth inhibition (CDI) is mediated by the CdiB/CdiA family of two-partner secretion proteins. CdiA effector proteins are exported onto the surface of CDI(+) inhibitor cells, where they interact with susceptible bacteria and deliver effectors/toxins derived from their C-terminal regions (CdiA-CT). CDI(+) cells also produce an immunity protein that binds the CdiA-CT and blocks its activity to prevent autoinhibition. Here, we show that the CdiA-CT from uropathogenic Escherichia coli strain 536 (UPEC536) is a latent tRNase that requires activation by the biosynthetic enzyme CysK (O-acetylserine sulfhydrylase A). UPEC536 CdiA-CT exhibits no nuclease activity in vitro, but cleaves within transfer RNA (tRNA) anti-codon loops when purified CysK is added. CysK and CdiA-CT form a stable complex, and their binding interaction appears to mimic that of the CysK/CysE cysteine synthase complex. CdiA-CT activation is also required for growth inhibition. Synthesis of CdiA-CT in E. coli cysK(+) cells arrests cell growth, whereas the growth of ΔcysK mutants is unaffected by the toxin. Moreover, E. coli ΔcysK cells are completely resistant to inhibitor cells expressing UPEC536 CdiA, indicating that CysK is required to activate the tRNase during CDI. Thus, CysK acts as a permissive factor for CDI, providing a potential mechanism to modulate growth inhibition in target cells.

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More information

Accepted/In Press date: 20 January 2012
Published date: February 2012

Identifiers

Local EPrints ID: 442657
URI: http://eprints.soton.ac.uk/id/eprint/442657
ISSN: 0890-9369
PURE UUID: 1a228b7f-8cfd-42de-b6e6-8fc165e0acf0
ORCID for Jeremy Webb: ORCID iD orcid.org/0000-0003-2068-8589

Catalogue record

Date deposited: 22 Jul 2020 16:31
Last modified: 18 Feb 2021 17:08

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Contributors

Author: Jeremy Webb ORCID iD
Author: Elie Diner
Author: Christina Beck
Author: David A. Low
Author: Christopher S. Hayes

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