Dr. Cezar Khursigara

Starting from a very young age, I have always had a strong interest in biology and the natural world around us. But it was a summer work-study program at Ryerson University where I first became fascinated with the world of bacteria. Under the guidance of Dr. Debora Foster, I studied how pathogenic bacteria such as enteropathogenic and enterohemorrhagic E. coli (EPEC and EHEC) caused disease in humans. This early experience opened my eyes to the world of biological research and showed me how exciting, stimulating and rewarding a laboratory environment could be. I then moved from Toronto to Montreal, where I obtained my Ph.D. from McGill University for my work elucidating the mechanisms of iron-uptake by Gram-negative bacteria, under the supervision of Dr. James Coulton. This experience in combining biochemical, biophysical and structural techniques to characterize protein-protein interactions laid the foundation for my drive to apply a multidisciplinary approach to study bacterial processes. In the course this work I realized how the visualization of molecular interactions using microscopy and high-resolution structural techniques could complement and greatly extend conclusions drawn from more traditional methods. This insight prompted me to move to the United States to work at the National Cancer Institute of the National Institutes of Health (NIH). As a Visiting Fellow in the laboratory of Dr. Sriram Subramaniam, I helped pioneer cutting-edge cryo-electron microscopy techniques alongside molecular and biochemical methods to investigate the molecular architecture of receptor complexes involved in directed bacterial movement, or chemotaxis. My laboratory continues to apply this multidisciplinary approach to the study of a variety of bacterial processes.

• B.Sc. Department of Chemistry and Biology, Ryerson University
• Ph.D. Department of Microbiology and Immunology, McGill University
• PDF. Laboratory of Cell Biology, National Cancer Institute, National Institutes of Health

Bacterial cell biology has seen a renaissance in the past several years that has been spurred in part by advances in imaging techniques. Major advances in fluorescent microscopy, cryo-electron microscopy (2-D) and cryo-electron tomography (3-D) have provided new insight into bacterial ultrastructures that accomplish fundamental processes, such as cell growth and movement. Advances in imaging are also providing evidence that these cellular systems and assemblies are not only highly complex, but generally function in concert to accomplish cellular goals.

The research in our laboratory focuses on elucidating the structure and function of protein complexes involved in complex biological processes. We are particularly interested in the macromolecular assemblies that govern bacterial cell division, cell-to-cell interaction, biofilm formation, motility and chemotaxis. Moreover, with the emergence of a growing number of multi-drug resistant strains of bacteria there is a pressing need to identify new drug targets. Accordingly, these essential bacterial processes provide a number of exciting candidates. My research group is taking a multidisciplinary approach to answer fundamental questions related to these essential cellular processes. By combining cryo-electron microscopy and tomography with biochemical, biophysical, molecular and cellular techniques, our goal is to identify potential therapeutics that can target a broad spectrum of disease-causing bacteria. We also seek to develop novel imaging techniques, including correlative methods using fluorescent and cryo-electron microscopy. We hope that the imaging methods we develop in this research program will transcend bacterial studies and significantly impact applications in diverse biological fields, thus leading to advances in structural biology, nanotechnology, ecology and medicine, among others.

We are currently accepting graduate students who share the lab's vision for understanding bacterial processes and developing cutting-edge imaging methods. Potential students with interests in cellular imaging and electron microscopy, microbiology, molecular and cellular biology, biochemistry, biophysics, and their applications to medically and environmentally relevant problems are encouraged to contact us.

2017

• The LPS Transport Pathway: A New Target for the Development of Gram-negative Antibiotics.
  Berezuk AM, Khursigara CM. 2017. In Antibiotic Drug Discovery: New Targets and Molecular Entities (Firestine S, Lister T, eds.) Royal Society of Chemistry, Cambridge, UK
  
• SLC6A14 Is a Genetic Modifier of Cystic Fibrosis That Regulates Pseudomonas aeruginosa Attachment to Human Bronchial Epithelial Cells.
  Di Paola M, Park AJ, Ahmadi S, Roach EJ, Wu YS, Struder-Kypke M, Lam JS, Bear CE, Khursigara CM.
  MBio. 2017 Dec 19;8(6).
  
• Potentiation of Tobramycin by Silver Nanoparticles against Pseudomonas aeruginosa Biofilms.
  Habash MB, Goodyear MC, Park AJ, Surette MD, Vis EC, Harris RJ, Khursigara CM.
  Antimicrob Agents Chemother. 2017 Oct 24;61(11).
  
• Bypassing the Need for the Transcriptional Activator EarA through a Spontaneous Deletion in the BRE Portion of the fla Operon Promoter in Methanococcus maripaludis.
  Ding Y, Berezuk A, Khursigara CM, Jarrell KF.
  Front Microbiol. 2017 Jul 17;8:1329.
  
• A Genotypic Analysis of Five P. aeruginosa Strains after Biofilm Infection by Phages Targeting Different Cell Surface Receptors.
  Pires DP, Dötsch A, Anderson EM, Hao Y, Khursigara CM, Lam JS, Sillankorva S, Azeredo J.
  Front Microbiol. 2017 Jun 30;8:1229.
  
• Phylogenetic distribution of the euryarchaeal archaellum regulator EarA and complementation of a Methanococcus maripaludis ∆earA mutant with heterologous earA homologues.
  Ding Y, Berezuk A, Khursigara CM, Jarrell KF.
  Microbiology. 2017 May 23.
  
• Complementation of an aglB Mutant of Methanococcus maripaludis with Heterologous Oligosaccharyltransferases.
  Ding Y, Vrionis HA, Schneider J, Berezuk A, Khursigara CM, Jarrell KF.
  PLoS One. 2016 Dec 1;11(12):e0167611.
  
• Identification of the First Transcriptional Activator of an Archaellum Operon in a Euryarchaeon.
  Ding Y, Nash J, Berezuk A, Khursigara CM, Langelaan DN, Smith SP, Jarrell KF.
  Mol Microbiol. 2016 Oct;102(1):54-70.

2016

• Roach EJ, Wroblewski C, Seidel L, Berezuk AM, Brewer D, Kimber MS, Khursigara CM. Structure and Mutational Analysis of Escherichia coli ZapD Reveals Charged Residues Involved in FtsZ Filament Bundling. J Bacteriol. (in press)
• Park AJ, Krieger JR, Khursigara CM. Survival proteomes: the emerging proteotype of antimicrobial resistance. FEMS Microbiol Rev. (in press)
• Ding Y, Lau Z, Logan SM, Kelly JF, Berezuk A, Khursigara CM, Jarrell KF. Effects of growth conditions on archaellation and N-glycosylation in Methanococcus maripaludis. Microbiology. 2016 Feb;162(2):339-50. 2015

2015

• Park AJ, Murphy K, Surette MD, Bandoro C, Krieger JR, Taylor P, Khursigara CM. Tracking the Dynamic Relationship between Cellular Systems and Extracellular Subproteomes in Pseudomonas aeruginosa Biofilms. J Proteome Res. 2015 Nov 6;14(11):4524-37.
• Rocker AJ, Weiss AR, Lam JS, Van Raay TJ, Khursigara CM. Visualizing and quantifying Pseudomonas aeruginosa infection in the hindbrain ventricle of zebrafish using confocal laser scanning microscopy. J Microbiol Methods. 2015 Oct;117:85-94.
• Hao Y, Murphy K, Lo RY, Khursigara CM, Lam JS. Single-Nucleotide Polymorphisms Found in the migA and wbpX Glycosyltransferase Genes Account for the Intrinsic Lipopolysaccharide Defects Exhibited by Pseudomonas aeruginosa PA14. J Bacteriol. 2015 Sep;197(17):2780-91.
• Ding Y, Uchida K, Aizawa S, Murphy K, Berezuk A, Khursigara CM, Chong JP, Jarrell KF. Effects of N-glycosylation site removal in archaellins on the assembly and function of archaella in Methanococcus maripaludis. PLoS One. 2015 Feb 20;10(2):e0116402.
• McDonald JA, Fuentes S, Schroeter K, Heikamp-deJong I, Khursigara CM, de Vos WM, Allen-Vercoe E. Simulating distal gut mucosal and luminal communities using packed-column biofilm reactors and an in vitro chemostat model. J Microbiol Methods. 2015 Jan;108:36-44.

2014

• Habash MB, Park AJ, Vis EC, Harris RJ, Khursigara CM. Synergy of Silver Nanoparticles and Aztreonam against Pseudomonas aeruginosa PAO1 Biofilms. Antimicrob Agents Chemother. 2014 Jul 21. pii: AAC.03170-14. (in press)
• Park AJ, Surette MD, Khursigara CM. Antimicrobial targets localize to the extracellular vesicle-associated proteome of Pseudomonas aeruginosa grown in a biofilm. Front Microbiol. 2014 Sept 3;5:464.
• Berezuk AM, Goodyear M, Khursigara CM. Site-directed fluorescence labeling reveals a revised N-terminal membrane topology and functional periplasmic residues in the Escherichia coli cell division protein FtsK. J Biol Chem. 2014 Aug 22;289(34):23287-301.
• Roach EJ, Kimber MS, Khursigara CM. Crystal structure and site-directed mutational analysis reveals key residues involved in Escherichia coli ZapA function. J Biol Chem. 2014 Aug 22;289(34):23276-86.
• Park AJ, Murphy K, Krieger JR, Brewer D, Taylor P, Habash M, Khursigara CM. A temporal examination of the planktonic and biofilm proteome of whole cell Pseudomonas aeruginosa PAO1 using quantitative mass spectrometry. Mol Cell Proteomics. 2014 Apr;13(4):1095-105.
• Murphy K, Park AJ, Hao Y, Brewer D, Lam JS, Khursigara CM. Influence of O polysaccharides on biofilm development and outer membrane vesicle biogenesis in Pseudomonas aeruginosa PAO1. J Bacteriol. 2014 Apr;196(7):1306-17.

2013

• McDonald JA, Schroeter K, Fuentes S, Heikamp-Dejong I, Khursigara CM, de Vos WM, Allen-Vercoe E. Evaluation of microbial community reproducibility, stability and composition in a human distal gut chemostat model. J Microbiol Methods. 2013 Nov;95(2):167-74.
• Eleftheriou NM, Ge X, Kolesnik J,Falconer SB, Harris RJ, Khursigara C, Brown ED, Brennan JD. Entrapment of Living Bacterial Cells in Low-Concentration Silica Materials Preserves Cell Division and Promoter Regulation. Chem Mater 2013, 25(23), pp 4798–4805.

2012

• Guri A, Griffiths M, Khursigara CM, Corredig M. The effect of milk fat globules on adherence and internalization of Salmonella Enteritidis to HT-29 cells. J Dairy Sci. 2012 Sep 26.

2011

• Khursigara CM, Lan G, Neumann S, Wu X, Ravindran S, Borgnia MJ, Sourjik V, Milne J, Tu Y, Subramaniam S. (2011) Lateral density of receptor arrays in the membrane plane influences sensitivity of the E. coli chemotaxis response.MBO J. May 4;30(9):1719-29.
• Li M, Khursigara CM, Subramaniam S, Hazelbauer GL. (2011) Chemotaxis kinase CheA is activated by three neighbouring chemoreceptor dimers as effectively as by receptor clusters. Mol Microbiol. Feb;79(3):677-85

2010 and earlier

• Khursigara CM, Wu X, Zhang P, Lefman J, Subramaniam S. (2008) Role of HAMP domains in transmembrane signaling by bacterial chemoreceptors. Proceedings of the National Academy of Sciences (USA). 105(43):16555-60.
• Khursigara CM, Wu X, Subramaniam S. (2008) Chemoreceptors in Caulobacter crescentus: trimers of receptor dimers in a partially ordered hexagonally packed array. J Bacteriol. 190(20):6805-10.
• *Zhang P, *Khursigara CM, Hartnell L, Subramaniam S. (2007) Direct visualization of E. coli chemotaxis receptor arrays using cryo-electron microscopy. Proceedings of the National Academy of Sciences (USA). 104(10):3777-3781. * equal contribution
• Pawelek PD, Croteau N, Ng-Thow-Hing C, Khursigara CM, Moiseeva N, Allaire M, Coulton JW. (2006) Structure of TonB in complex with FhuA, E. coli outer membrane receptor. Science. 312(5778):1399-402.
• Khursigara CM, De Crescenzo G, Pawelek, PD, Coulton, JW. (2004) Enhanced binding of TonB to a ligand-loaded outer membrane receptor: role of the oligomeric state of TonB in formation of a functional FhuA-TonB complex. J Biol Chem. 279(9):7405-12.
• Khursigara CM, De Crescenzo G, Pawelek PD, Coulton JW. (2005) Deletion of the proline-rich region of TonB disrupts formation of a 2:1 complex with FhuA, an outer membrane receptor of Escherichia coli. Protein Science. 14(5):1266-73.
• Khursigara CM, De Crescenzo G, Pawelek PD, Coulton JW. (2005) Kinetic analyses reveal multiple steps in forming TonB-FhuA complexes from Escherichia coli. Biochemistry. 4(9):3441-53.
• Khursigara C, Abul-Milh M, Lau B, Giron JA, Lingwood CA, Barnett Foster DE. (2001) Enteropathogenic Escherichia coli virulence factor bundle-forming pilus has a binding specificity for phosphatidylethanolamine. Infection and Immunity. 69(11):6573-9.