): M. pneumoniae is the leading cause of pneumonia in the age group that includes older children and young adults. This cell wall-less prokaryote is a member of a taxonomic class that includes numerous pathogens in humans, domestic animals, plants, and invertebrates. Despite their impact on public health and agriculture, mycoplasmas are poorly understood. Just as modern medicine is grounded in the detailed understanding of human anatomy, physiology, cell biology and gene regulation, more effective means of prevention, control and resolution of infections requires the elucidation of the basic biology and virulence factors of the organisms involved. Until recently, however, mycoplasmas have been refractory to the techniques of genetics and molecular biology that have contributed to major advances in defining the function and regulation of virulence factors in other important pathogens. Adherence of M. pneumoniae to the respiratory epithelium (cytadherence) is essential to successful colonization and disease. Cytadherence is a complex process involving adhesin proteins as well as accessory proteins required for the proper distribution and disposition of adhesins on the mycoplasma surface. The focus of this proposal is the detailed analysis of the operon that includes the genes for the adhesin, P30, and the cytadherence-accessory proteins, HMW1 and HMW3, as well as a cytadherence regulatory locus (crl) required for expression of HMW1 and HMW3. Nucleotide sequence analysis predicts that the genes for HMW1 and HMW3 are components of a multicistronic operon. The start site for transcription will be established by primer extension, and sequences required for expression will be evaluated using a reporter system and genetic complementation. Transcript levels will be compared by RNA slot blot hybridization. Transposon insertion sites in crl will be mapped, and excision deletion mutants isolated and characterized. Finally, the products of the other open reading frames in the hmw operon will be identified and their subcellular location established. Their requirement for the localization of HMW3 to, and assembly of, the attachment organelle will be assessed by immuno-electron microscopy of mutant strains.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI023362-14
Application #
2886507
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Taylor, Christopher E,
Project Start
1986-09-01
Project End
2000-04-30
Budget Start
1999-05-01
Budget End
2000-04-30
Support Year
14
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Georgia
Department
Microbiology/Immun/Virology
Type
Schools of Arts and Sciences
DUNS #
City
Athens
State
GA
Country
United States
Zip Code
30602
Hasselbring, Benjamin M; Sheppard, Edward S; Krause, Duncan C (2012) P65 truncation impacts P30 dynamics during Mycoplasma pneumoniae gliding. J Bacteriol 194:3000-7
Cloward, Jason M; Krause, Duncan C (2011) Loss of co-chaperone TopJ impacts adhesin P1 presentation and terminal organelle maturation in Mycoplasma pneumoniae. Mol Microbiol 81:528-39
Chang, How-Yi; Jordan, Jarrat L; Krause, Duncan C (2011) Domain analysis of protein P30 in Mycoplasma pneumoniae cytadherence and gliding motility. J Bacteriol 193:1726-33
Chang, How-Yi; Prince, Oliver A; Sheppard, Edward S et al. (2011) Processing is required for a fully functional protein P30 in Mycoplasma pneumoniae gliding and cytadherence. J Bacteriol 193:5841-6
Lai, Jen-Feng; Zindl, Carlene L; Duffy, Lynn B et al. (2010) Critical role of macrophages and their activation via MyD88-NF?B signaling in lung innate immunity to Mycoplasma pneumoniae. PLoS One 5:e14417
Cloward, Jason M; Krause, Duncan C (2010) Functional domain analysis of the Mycoplasma pneumoniae co-chaperone TopJ. Mol Microbiol 77:158-69
Cloward, Jason M; Krause, Duncan C (2009) Mycoplasma pneumoniae J-domain protein required for terminal organelle function. Mol Microbiol 71:1296-307
Bose, Stephanie R; Balish, Mitchell F; Krause, Duncan C (2009) Mycoplasma pneumoniae cytoskeletal protein HMW2 and the architecture of the terminal organelle. J Bacteriol 191:6741-8
Waldo 3rd, Robert H; Krause, Duncan C (2006) Synthesis, stability, and function of cytadhesin P1 and accessory protein B/C complex of Mycoplasma pneumoniae. J Bacteriol 188:569-75
Waldo 3rd, Robert H; Jordan, Jarrat L; Krause, Duncan C (2005) Identification and complementation of a mutation associated with loss of Mycoplasma pneumoniae virulence-specific proteins B and C. J Bacteriol 187:747-51

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