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Construction of a bacterial autoinducer detection system in mammalian cells

Biological Procedures Online volume 6, pages 268–276 (2004)Cite this article

Abstract

Quorum sensing (QS) is a cell density-dependent signaling system used by bacteria to coordinate gene expression within a population. QS systems in Gram negative bacteria consist of transcription factors of the LuxR family and their acyl homoserine lactone (AHL) ligands. We describe here a method for examining QS signaling systems in mammalian cells that uses engineered LuxR-type proteins from the opportunistic pathogen, Pseudomonas aeruginosa, which can function as AHL-dependent transcription factors. The engineered proteins respond to their cognate ligands and display sequence specific DNA binding properties. This system has several potential biotechnological and biological applications. It may be used to characterize any LuxR-type protein, screen animal and plant cell extracts or exudates for compounds that mimic or interfere with AHL signaling or to screen different cell types for AHL inactivating activities.

Abbreviations

QS:

quorum sensing

AHL:

acylhomoserine lactone

3O-C12-HSL:

N-3-oxodocanoyl-homoserine lactone

C4-HSL:

N-butyryl homoserine lactone

3O-C6-HSL:

N-3-oxoheptanoyl homoserine lactone

T3N:

protein module containing three copies of the VP16 transcriptional activation domain (TAD) and the nuclear localization signal (NLS) from SV40 T antigen

DAPI:

4′6′-diamidino-2-phenylindone

SMART:

Simple Modular Architecture Research Tool

AIBD:

autoinducer binding domain

HTH:

helix-turn-helix DNA binding domain

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Authors and Affiliations

  1. Department of Microbiology & Immunology, Texas Tech University Health Sciences Center, Lubbock, Texas

    Erin K. Shiner

  2. Department of Cell Biology & Biochemistry, Texas Tech University Health Sciences Center, Lubbock, Texas

    Sheila Reddy & Simon C. Williams

  3. Southwest Cancer Center at UMC, Lubbock, Texas

    Sheila Reddy & Simon C. Williams

  4. Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, Texas

    Cody Timmons & Guigen Li

  5. Department of Surgery, Texas Tech University Health Sciences Center, 3601 4th Street, 79430, Lubbock, Texas

    Kendra P. Rumbaugh Ph.D.

Authors
  1. Erin K. Shiner
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  2. Sheila Reddy
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  3. Cody Timmons
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  4. Guigen Li
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  5. Simon C. Williams
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  6. Kendra P. Rumbaugh Ph.D.
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Corresponding author

Correspondence to Kendra P. Rumbaugh Ph.D..

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Published: December 30, 2004.

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Shiner, E.K., Reddy, S., Timmons, C. et al. Construction of a bacterial autoinducer detection system in mammalian cells. Biol. Proced. Online 6, 268–276 (2004). https://doi.org/10.1251/bpo98

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  • DOI: https://doi.org/10.1251/bpo98

Indexing terms

  • Pseudomonas aeruginosa autoinducer
  • LuxR protein

Biological Procedures Online

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