Structural basis for virulence regulation in Vibrio cholerae by unsaturated fatty acid components of bile

Cruite, Justin T.; Kovacikova, Gabriela; Clark, Kenzie A.; Woodbrey, Anne K.; Skorupski, Karen; Kull, F. Jon

doi:10.1038/s42003-019-0686-x

Title: Structural basis for virulence regulation in Vibrio cholerae by unsaturated fatty acid components of bile

Journal Article · Thu Nov 28 00:00:00 EST 2019 · Communications Biology

DOI:https://doi.org/10.1038/s42003-019-0686-x· OSTI ID:1624534

Cruite, Justin T. ^[1]; Kovacikova, Gabriela ^[2];

^[3]; Woodbrey, Anne K. ^[4]; Skorupski, Karen ^[2]; Kull, F. Jon ^[5]

Geisel School of Medicine at Dartmouth College, Hanover, NH (United States). Department of Biochemistry and Cell Biology; Dartmouth College; Guarini School of Graduate and Advanced Studies, Hanover, NH (United States).
Geisel School of Medicine at Dartmouth College, Hanover, NH (United States). Department of Microbiology and Immunology
Dartmouth College, Hanover, NH (United States). Dept. of Chemistry
Dartmouth College; Guarini School of Graduate and Advanced Studies, Hanover, NH (United States); Dartmouth College, Hanover, NH (United States). Dept. of Chemistry
Geisel School of Medicine at Dartmouth College, Hanover, NH (United States). Department of Biochemistry and Cell Biology; Dartmouth College; Guarini School of Graduate and Advanced Studies, Hanover, NH (United States); Dartmouth College, Hanover, NH (United States). Dept. of Chemistry

The AraC/XylS-family transcriptional regulator ToxT is the master virulence activator of Vibrio cholerae, the gram-negative bacterial pathogen that causes the diarrheal disease cholera. Unsaturated fatty acids (UFAs) found in bile inhibit the activity of ToxT. Crystal structures of inhibited ToxT bound to UFA or synthetic inhibitors have been reported, but no structure of ToxT in an active conformation had been determined. Here we present the 2.5 Å structure of ToxT without an inhibitor. The structure suggests release of UFA or inhibitor leads to an increase in flexibility, allowing ToxT to adopt an active conformation that is able to dimerize and bind DNA. Small-angle X-ray scattering was used to validate a structural model of an open ToxT dimer bound to the cholera toxin promoter. The results presented here provide a detailed structural mechanism for virulence gene regulation in V. cholerae by the UFA components of bile and other synthetic ToxT inhibitors.

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Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 1624534

Journal Information:: Communications Biology, Vol. 2, Issue 1; ISSN 2399-3642

Publisher:: Springer NatureCopyright Statement

Country of Publication:: United States

Language:: English

References (40)

Evidence for Multimerization of Neu Proteins Involved in Polysialic Acid Synthesis in Escherichia coli K1 Using Improved LexA-Based Vectors Daines, D. A.; Silver, R. P. Journal of Bacteriology, Vol. 182, Issue 18 https://doi.org/10.1128/JB.182.18.5267-5270.2000	journal	September 2000
Effect of Fatty Acids and Cholesterol Present in Bile on Expression of Virulence Factors and Motility of Vibrio cholerae Chatterjee, Arpita; Dutta, Pradeep K.; Chowdhury, Rukhsana Infection and Immunity, Vol. 75, Issue 4 https://doi.org/10.1128/IAI.01435-06	journal	January 2007
Regulation of virulence in Vibrio cholerae : the ToxR regulon Childers, Brandon M.; Klose, Karl E. Future Microbiology, Vol. 2, Issue 3 https://doi.org/10.2217/17460913.2.3.335	journal	June 2007
A new LexA-based genetic system for monitoring and analyzing protein heterodimerization in Escherichia coli Dimitrova, M.; Younès-Cauet, G.; Oertel-Buchheit, P. Molecular and General Genetics MGG, Vol. 257, Issue 2 https://doi.org/10.1007/s004380050640	journal	January 1998
Arac/XylS family of transcriptional regulators. Gallegos, M. T.; Schleif, R.; Bairoch, A. Microbiology and molecular biology reviews : MMBR, Vol. 61, Issue 4 https://doi.org/10.1128/.61.4.393-410.1997	journal	January 1997
Vibrio cholerae ToxT Independently Activates the Divergently Transcribed aldA and tagA Genes Withey, Jeffrey H.; DiRita, Victor J. Journal of Bacteriology, Vol. 187, Issue 23 https://doi.org/10.1128/JB.187.23.7890-7900.2005	journal	December 2005
Pathogenic Potential of Environmental Vibrio cholerae Strains Carrying Genetic Variants of the Toxin-Coregulated Pilus Pathogenicity Island Faruque, Shah M.; Kamruzzaman, M.; Meraj, Ismail M. Infection and Immunity, Vol. 71, Issue 2 https://doi.org/10.1128/IAI.71.2.1020-1025.2003	journal	February 2003
Regulation of gene expression in Vibrio cholerae by ToxT involves both antirepression and RNA polymerase stimulation Yu, Rosa R.; DiRita, Victor J. Molecular Microbiology, Vol. 43, Issue 1 https://doi.org/10.1046/j.1365-2958.2002.02721.x	journal	January 2002
Identification and Characterization of the Functional Toxboxes in the Vibrio cholerae Cholera Toxin Promoter Dittmer, J. B.; Withey, J. H. Journal of Bacteriology, Vol. 194, Issue 19 https://doi.org/10.1128/JB.00952-12	journal	July 2012
Mechanism for Inhibition of Vibrio cholerae ToxT Activity by the Unsaturated Fatty Acid Components of Bile Plecha, Sarah C.; Withey, Jeffrey H. Journal of Bacteriology, Vol. 197, Issue 10 https://doi.org/10.1128/JB.02409-14	journal	March 2015
Mechanism of ToxT-Dependent Transcriptional Activation at the Vibrio cholerae tcpA Promoter Hulbert, Robin R.; Taylor, Ronald K. Journal of Bacteriology, Vol. 184, Issue 20 https://doi.org/10.1128/JB.184.20.5533-5544.2002	journal	October 2002
A new class of inhibitors of the AraC family virulence regulator Vibrio cholerae ToxT Woodbrey, Anne K.; Onyango, Evans O.; Pellegrini, Maria Scientific Reports, Vol. 7, Issue 1 https://doi.org/10.1038/srep45011	journal	March 2017
N-terminal Residues of the Vibrio cholerae Virulence Regulatory Protein ToxT Involved in Dimerization and Modulation by Fatty Acids Childers, Brandon M.; Cao, Xiaohang; Weber, Gregor G. Journal of Biological Chemistry, Vol. 286, Issue 32 https://doi.org/10.1074/jbc.M111.258780	journal	June 2011
The toxbox: specific DNA sequence requirements for activation of Vibrio cholerae virulence genes by ToxT Withey, Jeffrey H.; DiRita, Victor J. Molecular Microbiology, Vol. 59, Issue 6 https://doi.org/10.1111/j.1365-2958.2006.05053.x	journal	March 2006
A Modified ToxT Inhibitor Reduces Vibrio cholerae Virulence in Vivo Woodbrey, Anne K.; Onyango, Evans O.; Kovacikova, Gabriela Biochemistry, Vol. 57, Issue 38 https://doi.org/10.1021/acs.biochem.8b00667	journal	August 2018
PHENIX : building new software for automated crystallographic structure determination Adams, Paul D.; Grosse-Kunstleve, Ralf W.; Hung, Li-Wei Acta Crystallographica Section D Biological Crystallography, Vol. 58, Issue 11 https://doi.org/10.1107/S0907444902016657	journal	October 2002
Identification of Residues Critical for the Function of the Vibrio cholerae Virulence Regulator ToxT by Scanning Alanine Mutagenesis Childers, Brandon M.; Weber, Gregor G.; Prouty, Michael G. Journal of Molecular Biology, Vol. 367, Issue 5 https://doi.org/10.1016/j.jmb.2007.01.061	journal	April 2007
Accurate SAXS Profile Computation and its Assessment by Contrast Variation Experiments Schneidman-Duhovny, Dina; Hammel, Michal; Tainer, John A. Biophysical Journal, Vol. 105, Issue 4 https://doi.org/10.1016/j.bpj.2013.07.020	journal	August 2013
Scatter : software for the analysis of nano- and mesoscale small-angle scattering Förster, S.; Apostol, L.; Bras, W. Journal of Applied Crystallography, Vol. 43, Issue 3 https://doi.org/10.1107/S0021889810008289	journal	April 2010
Activation of both acfA and acfD transcription by Vibrio cholerae ToxT requires binding to two centrally located DNA sites in an inverted repeat conformation: Activation of acfA and acfD by ToxT Withey, Jeffrey H.; DiRita, Victor J. Molecular Microbiology, Vol. 56, Issue 4 https://doi.org/10.1111/j.1365-2958.2005.04589.x	journal	March 2005
Small-Molecule Inhibitor of Vibrio cholerae Virulence and Intestinal Colonization Hung, D. T. Science, Vol. 310, Issue 5748 https://doi.org/10.1126/science.1116739	journal	October 2005
Structure of Vibrio cholerae ToxT reveals a mechanism for fatty acid regulation of virulence genes Lowden, M. J.; Skorupski, K.; Pellegrini, M. Proceedings of the National Academy of Sciences, Vol. 107, Issue 7 https://doi.org/10.1073/pnas.0915021107	journal	February 2010
Conjugated Linoleic Acid Reduces Cholera Toxin Production In Vitro and In Vivo by Inhibiting Vibrio cholerae ToxT Activity Withey, Jeffrey H.; Nag, Dhrubajyoti; Plecha, Sarah C. Antimicrobial Agents and Chemotherapy, Vol. 59, Issue 12 https://doi.org/10.1128/AAC.01029-15	journal	September 2015
Regulatory cascade controls virulence in Vibrio cholerae. DiRita, V. J.; Parsot, C.; Jander, G. Proceedings of the National Academy of Sciences, Vol. 88, Issue 12 https://doi.org/10.1073/pnas.88.12.5403	journal	June 1991
*A branch in the ToxR regulatory cascade of Vibrio cholerae* revealed by characterization of toxT mutant strains** Champion, Gretchen A.; Neely, Melody N.; Brennan, Molly A. Molecular Microbiology, Vol. 23, Issue 2 https://doi.org/10.1046/j.1365-2958.1997.2191585.x	journal	January 1997
Virstatin inhibits dimerization of the transcriptional activator ToxT Shakhnovich, E. A.; Hung, D. T.; Pierson, E. Proceedings of the National Academy of Sciences, Vol. 104, Issue 7 https://doi.org/10.1073/pnas.0611643104	journal	February 2007
1.65 Å resolution structure of the AraC-family transcriptional activator ToxT from Vibrio cholerae Li, Jiaqin; Wehmeyer, Graham; Lovell, Scott Acta Crystallographica Section F Structural Biology Communications, Vol. 72, Issue 9 https://doi.org/10.1107/S2053230X1601298X	journal	August 2016
A novel DNA-binding motif in MarA: The first structure for an AraC family transcriptional activator Rhee, S.; Martin, R. G.; Rosner, J. L. Proceedings of the National Academy of Sciences, Vol. 95, Issue 18, p. 10413-10418 https://doi.org/10.1073/pnas.95.18.10413	journal	September 1998
Characterization of functional domains of the Vibrio cholerae virulence regulator ToxT: Structure and function of ToxT Prouty, Michael G.; Osorio, Carlos R.; Klose, Karl E. Molecular Microbiology, Vol. 58, Issue 4 https://doi.org/10.1111/j.1365-2958.2005.04897.x	journal	October 2005
Characterization of VPI Pathogenicity Island and CTXφ Prophage in Environmental Strains of Vibrio cholerae Mukhopadhyay, Asish K.; Chakraborty, Soumen; Takeda, Yoshifumi Journal of Bacteriology, Vol. 183, Issue 16 https://doi.org/10.1128/JB.183.16.4737-4746.2001	journal	August 2001
Structural Basis for Ligand-Regulated Oligomerization of AraC Soisson, S. M. Science, Vol. 276, Issue 5311 https://doi.org/10.1126/science.276.5311.421	journal	April 1997
Robust, high-throughput solution structural analyses by small angle X-ray scattering (SAXS) Hura, Greg L.; Menon, Angeli L.; Hammel, Michal Nature Methods, Vol. 6, Issue 8 https://doi.org/10.1038/nmeth.1353	journal	July 2009
Automatic processing of rotation diffraction data from crystals of initially unknown symmetry and cell constants Kabsch, W. Journal of Applied Crystallography, Vol. 26, Issue 6 https://doi.org/10.1107/S0021889893005588	journal	December 1993
Phaser crystallographic software McCoy, Airlie J.; Grosse-Kunstleve, Ralf W.; Adams, Paul D. Journal of Applied Crystallography, Vol. 40, Issue 4 https://doi.org/10.1107/S0021889807021206	journal	July 2007
The virulence gene activator ToxT from Vibrio cholerae is a member of the AraC family of transcriptional activators. Higgins, D. E.; Nazareno, E.; DiRita, V. J. Journal of Bacteriology, Vol. 174, Issue 21 https://doi.org/10.1128/jb.174.21.6974-6980.1992	journal	January 1992
Implementation and performance of SIBYLS: a dual endstation small-angle X-ray scattering and macromolecular crystallography beamline at the Advanced Light Source Classen, Scott; Hura, Greg L.; Holton, James M. Journal of Applied Crystallography, Vol. 46, Issue 1 https://doi.org/10.1107/s0021889812048698	journal	January 2013
Molecular mechanisms of virstatin resistance by non-O1/non-O139 strains of Vibrio cholerae Shakhnovich, Elizabeth A.; Sturtevant, Derek; Mekalanos, John J. Molecular Microbiology, Vol. 0, Issue 0 https://doi.org/10.1111/j.1365-2958.2007.05984.x	journal	November 2007
Environmental Signals Modulate ToxT-Dependent Virulence Factor Expression in Vibrio cholerae Schuhmacher, Darren A.; Klose, Karl E. Journal of Bacteriology, Vol. 181, Issue 5 https://doi.org/10.1128/jb.181.5.1508-1514.1999	journal	March 1999
Analysis of an Autoregulatory Loop Controlling ToxT, Cholera Toxin, and Toxin-Coregulated Pilus Production inVibrio cholerae Yu, Rosa R.; DiRita, Victor J. Journal of Bacteriology, Vol. 181, Issue 8 https://doi.org/10.1128/jb.181.8.2584-2592.1999	journal	April 1999
Growing Repertoire of AraC/XylS Activators Egan, Susan M. Journal of Bacteriology, Vol. 184, Issue 20 https://doi.org/10.1128/jb.184.20.5529-5532.2002	journal	October 2002

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