Abstract

Bacteroides fragilis is the leading cause of anaerobic bacteremia and intraabdominal abscesses. Studies have shown that the interaction of the B. fragilis capsular polysaccharide complex (CPC) with the host immune system leads to the formation of intraabdominal abscesses. The B. fragilis CPC is composed of at least five distinct capsular polysaccharides, polysaccharide A, B, C, D, and E (PS A-E). Each of these capsular polysaccharides is able to undergo a reversible ON/OFF phenotype known as phase variation. The biosynthesis loci containing the genes for the synthesis of PS A, PS B and PS C have been completely sequenced as well the region upstream of the PS D locus. These sequences have allowed for the analysis of the mechanism phase variation of these capsular polysaccharides at the genetic level. Preliminary data suggest that variation of the B. fragilis capsular polysaccharides occurs due to the DNA inversion of a small segment of DNA upstream of the respective polysaccharide biosynthesis locus. The invertible regions upstream of the PS A and PS D loci have been demonstrated to contain functional promoters. This application is divided into aims that will analyze phase variation of these capsular polysaccharides at the DNA, RNA and polysaccharide levels. Studies will be conducted to characterize the cis-acting elements necessary for DNA inversion of promoter regions and the trans-acting factors that mediate inversion. Additionally, the characteristics of phase variation of each of these capsular polysaccharides will be analyzed from organisms grown in vitro, and isolated from in vivo sites associated with health (colon) and disease (peritoneal cavity and abscesses) Due to the novelty of the phase variation of five distinct capsular polysaccharide by a single organism, we have the unique opportunity to determine if there is coordinate regulation between the expression of distinct capsular polysaccharides, and the role phase variation of the B. fragilis capsular polysaccharides may play in health and disease.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI044193-06
Application #
6698862
Study Section
Bacteriology and Mycology Subcommittee 2 (BM)
Program Officer
Korpela, Jukka K
Project Start
1998-02-01
Project End
2007-01-31
Budget Start
2004-02-01
Budget End
2005-01-31
Support Year
6
Fiscal Year
2004
Total Cost
$299,100
Indirect Cost

  Institution

Name
Brigham and Women's Hospital
Department
Type
DUNS #
030811269
City
Boston
State
MA
Country
United States
Zip Code
02115

  Publications

Wieland Brown, Laura C; Penaranda, Cristina; Kashyap, Purna C et al. (2013) Production of ?-galactosylceramide by a prominent member of the human gut microbiota. PLoS Biol 11:e1001610
Chatzidaki-Livanis, Maria; Weinacht, Katja G; Comstock, Laurie E (2010) Trans locus inhibitors limit concomitant polysaccharide synthesis in the human gut symbiont Bacteroides fragilis. Proc Natl Acad Sci U S A 107:11976-80
Chatzidaki-Livanis, Maria; Coyne, Michael J; Comstock, Laurie E (2009) A family of transcriptional antitermination factors necessary for synthesis of the capsular polysaccharides of Bacteroides fragilis. J Bacteriol 191:7288-95
Coyne, Michael J; Comstock, Laurie E (2008) Niche-specific features of the intestinal bacteroidales. J Bacteriol 190:736-42
Chatzidaki-Livanis, Maria; Coyne, Michael J; Roche-Hakansson, Hazeline et al. (2008) Expression of a uniquely regulated extracellular polysaccharide confers a large-capsule phenotype to Bacteroides fragilis. J Bacteriol 190:1020-6
Coyne, Michael J; Chatzidaki-Livanis, Maria; Paoletti, Lawrence C et al. (2008) Role of glycan synthesis in colonization of the mammalian gut by the bacterial symbiont Bacteroides fragilis. Proc Natl Acad Sci U S A 105:13099-104
Roche-Hakansson, Hazeline; Chatzidaki-Livanis, Maria; Coyne, Michael J et al. (2007) Bacteroides fragilis synthesizes a DNA invertase affecting both a local and a distant region. J Bacteriol 189:2119-24
Coyne, Michael J; Reinap, Barbara; Lee, Martin M et al. (2005) Human symbionts use a host-like pathway for surface fucosylation. Science 307:1778-81
Weinacht, Katja G; Roche, Hazeline; Krinos, Corinna M et al. (2004) Tyrosine site-specific recombinases mediate DNA inversions affecting the expression of outer surface proteins of Bacteroides fragilis. Mol Microbiol 53:1319-30
Coyne, Michael J; Weinacht, Katja G; Krinos, Corinna M et al. (2003) Mpi recombinase globally modulates the surface architecture of a human commensal bacterium. Proc Natl Acad Sci U S A 100:10446-51

Showing the most recent 10 out of 16 publications