Gaussia Luciferase as a Reporter for Quorum Sensing in Staphylococcus aureus

Gaussia luciferase (GLuc) is a secreted protein with significant potential for use as a reporter of gene expression in bacterial pathogenicity studies. To date there are relatively few examples of its use in bacteriology. In this study we show that GLuc can be functionally expressed in the human pathogen Staphylococcus aureus and furthermore show that it can be used as a biosensor for the agr quorum sensing (QS) system which employs autoinducing peptides to control virulence. GLuc was linked to the P3 promoter of the S. aureus agr operon. Biosensor strains were validated by evaluation of chemical agent-mediated activation and inhibition of agr. Use of GLuc enabled quantitative assessment of agr activity. This demonstrates the utility of Gaussia luciferase for in vitro monitoring of agr activation and inhibition.

[1]  R. Novick.  Autoinduction and signal transduction in the regulation of staphylococcal virulence , 2003, Molecular microbiology.

[2]  S. McLeod,et al.  Secreted Gaussia princeps Luciferase as a Reporter of Escherichia coli Replication in a Mouse Tissue Cage Model of Infection , 2014, PloS one.

[3]  B. Tannous,et al.  Gaussia luciferase reporter assay for monitoring biological processes in culture and in vivo , 2009, Nature Protocols.

[4]  F. Vandenesch,et al.  The agr P2 operon: an autocatalytic sensory transduction system in Staphylococcus aureus. , 1995, Molecular & general genetics : MGG.

[5]  M. Yohda,et al.  Biophysical characterization of highly active recombinant Gaussia luciferase expressed in Escherichia coli. , 2010, Biochimica et biophysica acta.

[6]  Kenzo Nishiguchi,et al.  High-Throughput Screening of Inhibitors Targeting Agr/Fsr Quorum Sensing in Staphylococcus aureus and Enterococcus faecalis , 2013, Bioscience, biotechnology, and biochemistry.

[7]  Min Xu,et al.  Transforming the Untransformable: Application of Direct Transformation To Manipulate Genetically Staphylococcus aureus and Staphylococcus epidermidis , 2012, mBio.

[8]  R. Novick.  Properties of a cryptic high-frequency transducing phage in Staphylococcus aureus. , 1967, Virology.

[9]  Brock F. Binkowski,et al.  Engineered Luciferase Reporter from a Deep Sea Shrimp Utilizing a Novel Imidazopyrazinone Substrate , 2012, ACS chemical biology.

[10]  A. Horswill,et al.  Impact of Environmental Cues on Staphylococcal Quorum Sensing and Biofilm Development* , 2016, The Journal of Biological Chemistry.

[11]  Bruce S. Edwards,et al.  Selective Chemical Inhibition of agr Quorum Sensing in Staphylococcus aureus Promotes Host Defense with Minimal Impact on Resistance , 2014, PLoS pathogens.

[12]  R. Novick,et al.  Transient interference with staphylococcal quorum sensing blocks abscess formation. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[13]  W. Chan,et al.  Targeting Staphylococcus aureus Quorum Sensing with Nonpeptidic Small Molecule Inhibitors , 2014, Journal of medicinal chemistry.

[14]  M. Otto.  Quorum-sensing control in Staphylococci -- a target for antimicrobial drug therapy? , 2004, FEMS microbiology letters.

[15]  C. Malone,et al.  Signal Biosynthesis Inhibition with Ambuic Acid as a Strategy To Target Antibiotic-Resistant Infections , 2017, Antimicrobial Agents and Chemotherapy.

[16]  A. Clatworthy,et al.  Targeting virulence: a new paradigm for antimicrobial therapy , 2007, Nature Chemical Biology.

[17]  D. Sturdevant,et al.  RNAIII-independent target gene control by the agr quorum-sensing system: insight into the evolution of virulence regulation in Staphylococcus aureus. , 2008, Molecular cell.

[18]  G. Asokan,et al.  WHO Global Priority Pathogens List: A Bibliometric Analysis of Medline-PubMed for Knowledge Mobilization to Infection Prevention and Control Practices in Bahrain , 2019, Oman medical journal.

[19]  W. Chan,et al.  Structure, activity and evolution of the group I thiolactone peptide quorum‐sensing system of Staphylococcus aureus , 2001, Molecular microbiology.

[20]  K. Winzer,et al.  Differential recognition of Staphylococcus aureus quorum-sensing signals depends on both extracellular loops 1 and 2 of the transmembrane sensor AgrC. , 2008, Journal of molecular biology.

[21]  Ralph Weissleder,et al.  A secreted luciferase for ex vivo monitoring of in vivo processes , 2008, Nature Methods.

[22]  Kenneth W. Bayles,et al.  A Genetic Resource for Rapid and Comprehensive Phenotype Screening of Nonessential Staphylococcus aureus Genes , 2013, mBio.

[23]  S. McAllister,et al.  Pulsed-Field Gel Electrophoresis Typing of Oxacillin-Resistant Staphylococcus aureus Isolates from the United States: Establishing a National Database , 2003, Journal of Clinical Microbiology.

[24]  W. Chan,et al.  Attenuating Staphylococcus aureus Virulence Gene Regulation: A Medicinal Chemistry Perspective , 2013, Journal of medicinal chemistry.

[25]  Rustam I. Aminov,et al.  A Brief History of the Antibiotic Era: Lessons Learned and Challenges for the Future , 2010, Front. Microbio..

[26]  Lee-Ann Tjon-Kon-Fat,et al.  Gaussia luciferase variant for high-throughput functional screening applications. , 2009, Analytical chemistry.

[27]  B. Bonev,et al.  Timing is everything - impact of naturally occurring Staphylococcus aureus AgrC cytoplasmic domain adaptive mutations on auto-induction. , 2019, Journal of bacteriology.

[28]  M. Stefanidou,et al.  Alternative Luciferase for Monitoring Bacterial Cells under Adverse Conditions , 2005, Applied and Environmental Microbiology.

[29]  T. Muir,et al.  Structure-activity analysis of synthetic autoinducing thiolactone peptides from Staphylococcus aureus responsible for virulence. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Roman G. Gerlach,et al.  Gaussia princeps Luciferase as a Reporter for Transcriptional Activity, Protein Secretion, and Protein-Protein Interactions in Salmonella enterica Serovar Typhimurium , 2011, Applied and Environmental Microbiology.

[31]  Philip J Hill,et al.  Construction and evaluation of multisite recombinatorial (Gateway) cloning vectors for Gram-positive bacteria , 2007, BMC Molecular Biology.

[32]  M. Lei,et al.  A single copy integration vector that integrates at an engineered site on the Staphylococcus aureus chromosome , 2012, BMC Research Notes.

[33]  Sai Bi,et al.  Chemiluminescence and Bioluminescence Imaging for Biosensing and Therapy: In Vitro and In Vivo Perspectives , 2019, Theranostics.

[34]  H. Ingmer,et al.  The agr Inhibitors Solonamide B and Analogues Alter Immune Responses to Staphylococccus aureus but Do Not Exhibit Adverse Effects on Immune Cell Functions , 2016, PloS one.

[35]  H. Raja,et al.  ω-Hydroxyemodin Limits Staphylococcus aureus Quorum Sensing-Mediated Pathogenesis and Inflammation , 2015, Antimicrobial Agents and Chemotherapy.

[36]  D. Picard,et al.  Secreted dual reporter assay with Gaussia luciferase and the red fluorescent protein mCherry , 2017, PloS one.

[37]  Yutaka Kuroda,et al.  Bacterial expression and re-engineering of Gaussia princeps luciferase and its use as a reporter protein. , 2015, Biochimica et biophysica acta.

[38]  Philip Hill,et al.  N-Acylhomoserine Lactones Antagonize Virulence Gene Expression and Quorum Sensing in Staphylococcus aureus , 2006, Infection and Immunity.

[39]  A. Sonenshein,et al.  CodY-Mediated Regulation of the Staphylococcus aureus Agr System Integrates Nutritional and Population Density Signals , 2014, Journal of bacteriology.

[40]  Blaise R. Boles,et al.  agr-Mediated Dispersal of Staphylococcus aureus Biofilms , 2008, PLoS pathogens.

[41]  Weibo Cai,et al.  NanoLuc: A Small Luciferase Is Brightening Up the Field of Bioluminescence. , 2016, Bioconjugate chemistry.

[42]  F. DeLeo,et al.  Vancomycin Resistance in Staphylococcus aureus
 , 2017, The Yale journal of biology and medicine.

[43]  Brian Gray,et al.  Targeting agr- and agr-Like Quorum Sensing Systems for Development of Common Therapeutics to Treat Multiple Gram-Positive Bacterial Infections , 2013, Sensors.

[44]  S. Projan,et al.  Bacterial virulence as a target for antimicrobial chemotherapy. , 2000, Current opinion in biotechnology.