This renewal aims to characterize structure-function relationships for type IV pili fibers, which key virulence factors for pathogenic Gram-negative bacteria. Structural analyses for type IV pilin subunits will be integrated with electron microscopy (EM), fiber diffraction, and small angle x-ray scattering (SAXS) structures of native fibers via objective Fourier correlation methods. These proposed studies, which span atomic to subcellular resolutions, will focus upon type IV pili from Neisseria gonorrhoeae, the causative agent of gonorrhea. Successful methods and results on gonococcal pili will allow complementary structural and mutational studies on pili from Pseudomonas aeruginosa, the causative agent on deadly opportunistic nosocomial infections, and Vibrio cholerae, the causative agent of cholera, to define conserved and variable aspects of type IV pili. Key questions concerning pilus structure-function relationships will be addressed including whether the N. gonorrhoeae pilin fold is representative of all type IV pilins, how extreme antigenic variation avoids disrupting the pilin fold and fiber assembly, the nature and significance of post-translational modifications, structural changes associated with fiber formation, species-specific conservation of surface regions acting in target cell recognition and accessory protein binding, the structural chemistry controlling bundling, structural characteristics of immunodominant regions, and optimal approaches to the design of cross- species vaccines. Structural results and hypotheses will be experimentally tested by quantitative correlations among diffraction and electron microscopy results and by mutational analyses. The proposed integrated multi-disciplinary studies provide innovation in determining challenging fiber-forming protein structures and in bridging the huge resolution gap between protein crystal structures and EM image reconstructions of subcellular organelles. Overall, these structural and mutational results will promote integration of ongoing biochemical, immunobiological, genetic, and functional studies to decipher the structural chemistry governing pilus actions in pathogenicity: host cell surface attachment, twitching motility, bacteriophage absorption, modulation of transformation efficiency and toleration of extreme sequence variability while retaining structural integrity and flexibility. This understanding of pilus structure-function relationships has long-term potential applications for drug and vaccine design against major bacterial diseases now showing increasing antibiotic resistance and threats to public health.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI022160-12A1
Application #
6046054
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Quackenbush, Robert L
Project Start
1985-07-01
Project End
2005-01-31
Budget Start
2000-02-01
Budget End
2001-01-31
Support Year
12
Fiscal Year
2000
Total Cost
$303,323
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Banerjee, Ankan; Tsai, Chi-Lin; Chaudhury, Paushali et al. (2015) FlaF Is a ?-Sandwich Protein that Anchors the Archaellum in the Archaeal Cell Envelope by Binding the S-Layer Protein. Structure 23:863-872
Shin, David S; Pratt, Ashley J; Tainer, John A (2014) Archaeal genome guardians give insights into eukaryotic DNA replication and damage response proteins. Archaea 2014:206735
Reindl, Sophia; Ghosh, Abhrajyoti; Williams, Gareth J et al. (2013) Insights into FlaI functions in archaeal motor assembly and motility from structures, conformations, and genetics. Mol Cell 49:1069-82
Yamagata, Atsushi; Milgotina, Ekaterina; Scanlon, Karen et al. (2012) Structure of an essential type IV pilus biogenesis protein provides insights into pilus and type II secretion systems. J Mol Biol 419:110-24
Yannone, Steven M; Hartung, Sophia; Menon, Angeli L et al. (2012) Metals in biology: defining metalloproteomes. Curr Opin Biotechnol 23:89-95
Li, Juliana; Egelman, Edward H; Craig, Lisa (2012) Structure of the Vibrio cholerae Type IVb Pilus and stability comparison with the Neisseria gonorrhoeae type IVa pilus. J Mol Biol 418:47-64
Hartung, Sophia; Arvai, Andrew S; Wood, Timothy et al. (2011) Ultrahigh resolution and full-length pilin structures with insights for filament assembly, pathogenic functions, and vaccine potential. J Biol Chem 286:44254-65
Ghosh, Abhrajyoti; Hartung, Sophia; van der Does, Chris et al. (2011) Archaeal flagellar ATPase motor shows ATP-dependent hexameric assembly and activity stimulation by specific lipid binding. Biochem J 437:43-52
Lim, Mindy S; Ng, Dixon; Zong, ZuSheng et al. (2010) Vibrio cholerae El Tor TcpA crystal structure and mechanism for pilus-mediated microcolony formation. Mol Microbiol 77:755-70
Burke, John E; Babakhani, Arneh; Gorfe, Alemayehu A et al. (2009) Location of inhibitors bound to group IVA phospholipase A2 determined by molecular dynamics and deuterium exchange mass spectrometry. J Am Chem Soc 131:8083-91

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