The goal of this project is to better understand the structure and function of a gonococcal membrane protein implicated in pathogenesis. Protein I (P.I) is a major outer membrane protein, and has been implicated in susceptibility to killing by serum and by polymorphonuclear neutrophils, and in ability to invade host cells. This application proposes to use molecular genetic techniques including transposon mutagenesis of cloned genes, and construction of strains carrying alternative forms of P.I. To provide its role in pathogenesis. Isogenic strains will be constructed containing either P.IA or P.IB or chimeric P.IA/P.IB hybrids, by transformation of competent gonococci with cloned P.I structural genes closely linked to a gene from chloramphenicol acetyl transferase. Attempts will be made to isolate P.I-mutants as well. These isogenic strains will be used in studies of serum killing, neutrophil susceptibility, and invasion of HL60 and primary human endometrial epithelial cell cultures, with or without addition of specific monoclonal and polyclonal sera directed at portions of these proteins. Epitopes on P.I will be mapped by analysis of the structure of chimeric P.IA/P.IB proteins, and by construction of recombinant fusion proteins. Previously describe genes for serum resistance and antibiotic resistance that are linked to the genes for P.I will be isolated by """"""""chromosome walking"""""""" from the cloned P.I genes. These studies will expand significantly our knowledge of the roles played in pathogenesis of P.I and will contribute towards rational vaccine development.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
4R37AI026837-06
Application #
3481333
Study Section
Special Emphasis Panel (NSS)
Project Start
1988-07-01
Project End
1998-06-30
Budget Start
1993-07-01
Budget End
1994-06-30
Support Year
6
Fiscal Year
1993
Total Cost
Indirect Cost
Name
University of North Carolina Chapel Hill
Department
Type
Schools of Medicine
DUNS #
078861598
City
Chapel Hill
State
NC
Country
United States
Zip Code
27599
Anderson, James E; Hobbs, Marcia M; Biswas, Gour D et al. (2003) Opposing selective forces for expression of the gonococcal lactoferrin receptor. Mol Microbiol 48:1325-37
Chen, Ching-Ju; Mclean, Dalton; Thomas, Christopher E et al. (2002) Point mutations in HpuB enable gonococcal HpuA deletion mutants to grow on hemoglobin. J Bacteriol 184:420-6
Turner, P C; Thomas, C E; Stojiljkovic, I et al. (2001) Neisserial TonB-dependent outer-membrane proteins: detection, regulation and distribution of three putative candidates identified from the genome sequences. Microbiology 147:1277-90
Cornelissen, C N; Anderson, J E; Boulton, I C et al. (2000) Antigenic and sequence diversity in gonococcal transferrin-binding protein A. Infect Immun 68:4725-35
Carson, S D; Stone, B; Beucher, M et al. (2000) Phase variation of the gonococcal siderophore receptor FetA. Mol Microbiol 36:585-93
Biswas, G D; Anderson, J E; Chen, C J et al. (1999) Identification and functional characterization of the Neisseria gonorrhoeae lbpB gene product. Infect Immun 67:455-9
Carson, S D; Klebba, P E; Newton, S M et al. (1999) Ferric enterobactin binding and utilization by Neisseria gonorrhoeae. J Bacteriol 181:2895-901
Chen, C J; Elkins, C; Sparling, P F (1998) Phase variation of hemoglobin utilization in Neisseria gonorrhoeae. Infect Immun 66:987-93
Turner, P C; Thomas, C E; Elkins, C et al. (1998) Neisseria gonorrhoeae heme biosynthetic mutants utilize heme and hemoglobin as a heme source but fail to grow within epithelial cells. Infect Immun 66:5215-23
Cornelissen, C N; Anderson, J E; Sparling, P F (1997) Characterization of the diversity and the transferrin-binding domain of gonococcal transferrin-binding protein 2. Infect Immun 65:822-8

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