In previous studies we have shown that the B. burgdorferi bmp chromosomal gene cluster encoding the paralogous Bmp membrane lipoproteins is present in all B. burgdorferi sensu lato. This cluster forms two complex transcriptional units that are transcribed into variable monocistronic and polycistronic messages to generate different bmp mRNA and gene product concentrations. Nutritional stress in Borrelia burgdorferi triggers the stringent response that modulates expression of many genes and is mediated by the alarmon (p)ppGpp. Our work now indicates that B. burgdorferi has a transcriptionally active rel/Bbu, gene needed for generation of (p)ppGpp and that the presence of (p)ppGpp is associated with the modulation of expression of bmp and other genes under different environmental conditions. The genetic, transcriptional, and biological characteristics of the bmp gene cluster makes it an ideal model to study chromosomal gene regulation both by trans regulators such as (p)ppGpp and sigma factors and by cis DNA sequences. The hypothesis underlying this project is that expression of bmp and other B burgdorferi genes is modulated in trans by the stringent response both directly by (p)ppGpp and indirectly through sigma 54 and sigma S and in cis by the DNA sequences of their 5' upstream promoter regions.
The specific aims of this project are: 1) characterize the regulation of rel/Bbu expression and the Rel/Bbu-mediated stringent response on bmp expression during growth of B. burgdorferi in BSKH media, in co-culture with tick cells, and in rat peritoneal culture chambers and ticks; 2) ascertain the roles of sigma S and sigma 54 in modulating bmp expression by the stringent response and their hierarchical interactions in B. burgdorferi in vitro and in vivo using bacterial genetics methods, DNA microarrays and proteomics; and 3) identify putative regulatory proteins binding to 5' upstream DNA sequences of the rpsL, bmpD, bmpC, bmpA and sigma S genes and their DNA binding sites. The proposed experiments will permit us to identify global factors, hierarchical regulatory networks and cis acting DNA sequences involved in the regulation of bmp genes, and will take studies of gene expression and modulation in B. burgdorferi from a descriptive paradigm to a paradigm framed by mechanistic and causal interpretations.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI043063-05A1
Application #
6617152
Study Section
Special Emphasis Panel (ZRG1-BM-1 (02))
Program Officer
Baker, Phillip J
Project Start
1998-07-01
Project End
2008-02-29
Budget Start
2003-03-01
Budget End
2004-02-29
Support Year
5
Fiscal Year
2003
Total Cost
$161,027
Indirect Cost
Name
New York Medical College
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
041907486
City
Valhalla
State
NY
Country
United States
Zip Code
10595
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Bryksin, Anton V; Tomova, Alexandra; Godfrey, Henry P et al. (2010) BmpA is a surface-exposed outer-membrane protein of Borrelia burgdorferi. FEMS Microbiol Lett 309:77-83
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Bryksin, Anton V; Godfrey, Henry P; Carbonaro, Carol A et al. (2005) Borrelia burgdorferi BmpA, BmpB, and BmpD proteins are expressed in human infection and contribute to P39 immunoblot reactivity in patients with Lyme disease. Clin Diagn Lab Immunol 12:935-40
Morozova, O V; Cabello, F C; Dobrotvorsky, A K (2004) Semi-nested PCR detection of Bartonella henselae in Ixodes persulcatus ticks from Western Siberia, Russia. Vector Borne Zoonotic Dis 4:306-9
Morozova, Olga V; Dobrotvorsky, Andrey K; Livanova, Natalya N et al. (2002) PCR detection of Borrelia burgdorferi sensu lato, tick-borne encephalitis virus, and the human granulocytic ehrlichiosis agent in Ixodes persulcatus ticks from Western Siberia, Russia. J Clin Microbiol 40:3802-4
Bugrysheva, Julia; Dobrikova, Elena Y; Godfrey, Henry P et al. (2002) Modulation of Borrelia burgdorferi stringent response and gene expression during extracellular growth with tick cells. Infect Immun 70:3061-7
Dobrikova, E Y; Bugrysheva, J; Cabello, F C (2001) Two independent transcriptional units control the complex and simultaneous expression of the bmp paralogous chromosomal gene family in Borrelia burgdorferi. Mol Microbiol 39:370-8
Gorbacheva, V Y; Godfrey, H P; Cabello, F C (2000) Analysis of the bmp gene family in Borrelia burgdorferi sensu lato. J Bacteriol 182:2037-42

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