Many microorganisms produce pili or firmbriae on their cell surface. The proteins vary in functon depending on the organism. In many cases these surface structures enable the bacterium to adhere to their host or organism they subsequently interact with. Thus these surface structures play a primary role in host-parasite interaction. The pili produced by Neisseria gonorrhoeae are no exception. These pili function in the attachment of the gonococci to the human host and therefore is a primary determinant of pathogenicity, a major factor in cell-cell interaction. The gene(s) encoding pilus synthesis is turned on and off at high frequency. The phenotypes (P+ and P-) can be determined easily by light microscopy. There are at least 50 pili serotypes. The heterogeneity lies at the carboxyterminal end; the amino terminal end for the pilus subunit is very similar if not identical. Very little is known about the genetic regulation of pili expression in N. gonorrhoeae. For these reasons, the pili of the gonococci represent an interesting system for studying the regulation of gene expression. We propose to study the basis for pilus expression in N. gonorrhoeae. Using techniques in genetics (mutagenesis and complementation), DNA biochemistry (recombinant cloning, DNA sequencing, Southern hybridization), protein chemistry (SDS PAGE, 2-dimensional gels, Western blotting) and immunology (immunoprecipitation) we hope to elucidate the mechanism underlying the heterogeneity of pilus type as well as the genetic and molecular mechanisms regulating the expression of pili synthesis by the gonococcus.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI020845-08
Application #
3130630
Study Section
Microbial Physiology and Genetics Subcommittee 2 (MBC)
Project Start
1983-06-15
Project End
1991-05-31
Budget Start
1990-06-01
Budget End
1991-05-31
Support Year
8
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Arvidson, C G; So, M (1995) The Neisseria transcriptional regulator PilA has a GTPase activity. J Biol Chem 270:26045-8
Arvidson, C G; So, M (1995) Interaction of the Neisseria gonorrhoeae PilA protein with the pilE promoter involves multiple sites on the DNA. J Bacteriol 177:2497-504
Waldbeser, L S; Ajioka, R S; Merz, A J et al. (1994) The opaH locus of Neisseria gonorrhoeae MS11A is involved in epithelial cell invasion. Mol Microbiol 13:919-28
Boyle-Vavra, S; So, M; Seifert, H S (1993) Transcriptional control of gonococcal pilE expression: involvement of an alternate sigma factor. Gene 137:233-6
Stein, D C; Chien, R; Seifert, H S (1992) Construction of a Neisseria gonorrhoeae MS11 derivative deficient in NgoMI restriction and modification. J Bacteriol 174:4899-906
Taha, M K; Dupuy, B; Saurin, W et al. (1991) Control of pilus expression in Neisseria gonorrhoeae as an original system in the family of two-component regulators. Mol Microbiol 5:137-48
Seifert, H S; Ajioka, R S; Paruchuri, D et al. (1990) Shuttle mutagenesis of Neisseria gonorrhoeae: pilin null mutations lower DNA transformation competence. J Bacteriol 172:40-6
Paruchuri, D K; Seifert, H S; Ajioka, R S et al. (1990) Identification and characterization of a Neisseria gonorrhoeae gene encoding a glycolipid-binding adhesin. Proc Natl Acad Sci U S A 87:333-7
Parge, H E; Bernstein, S L; Deal, C D et al. (1990) Biochemical purification and crystallographic characterization of the fiber-forming protein pilin from Neisseria gonorrhoeae. J Biol Chem 265:2278-85
Taha, M K; So, M; Seifert, H S et al. (1988) Pilin expression in Neisseria gonorrhoeae is under both positive and negative transcriptional control. EMBO J 7:4367-78

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