The genus Rickettsia contains important pathogens and consists of small gram-negative bacteria that replicate within eukaryotic host cells. Many Rickettsia species are associated with blood-feeding arthropods (ticks, lice, fleas and mites) that serve as vectors for their transmission to vertebrate hosts. Various Rickettsia species are the etiologic agents of typhus and spotted fevers in humans but others are apparently confined to arthropod hosts as endosymbionts. Members of the Rickettsia are the closest known microbial relatives of mitochondria with which they share the characteristics of highly reduced genomes and intimate metabolic interactions with their host cells. Historically, the slow and obligate intracellular growth of rickettsiae and their resistance to genetic manipulation presented substantial technical challenges to their study. Our laboratory has been at the forefront of both the recently developed capability to genetically manipulate rickettsiae and the discovery of rickettsial plasmids, which are extra-chromosomal autonomously replicating genetic elements. Plasmids in pathogenic bacteria are often associated with virulence functions involving particular metabolic capabilities or antibiotic resistance and are an important source of genetic diversity because they may be mobile, or genetically transmissible, via conjugation between bacteria of the same or different species. We hypothesize that plasmids are involved in rickettsial host adaptation/virulence and genetic plasticity. We propose a comprehensive analysis of the distribution, phylogeny and role of plasmids in the biology of rickettsiae associated with blood-feeding arthropods. We will use pulsed field gel electrophoresis and Southern blot techniques to define the distribution of plasmids within the genus. We will physically isolate and clone plasmids from Rickettsia species that range from non-pathogenic arthropod endosymbionts to human pathogens. The cloned plasmids will be sequenced with high throughput pyro-sequencing technology (454 Life Science Corp.) and subjected to phylogenetic analysis to determine the probable origin(s) and evolutionary history of plasmids within the genus, and obtain clues to their potential host adaptive functions. We will test our hypotheses that plasmids confer host adaptive functions and genetic diversity within rickettsiae through plasmid curing and phenotype analyses and plasmid conjugation experiments. The proposed research will delineate the distribution of plasmids and test their role in host adaptation/virulence in a group of medically important obligate intracellular bacteria that, until recently, were not believed to harbor plasmids.

Public Health Relevance

Obligate intracellular bacteria of the genus Rickettsia are transmitted by blood-feeding insects and ticks. Members of the Rickettsia cause disease in humans that includes typhus and spotted fevers. The proposed research will characterize newly discovered genetic components (plasmids) of the Rickettsia and how they may contribute to the ability of these bacteria to infect insects, ticks and humans. The results of this research may provide new tools to control Rickettsia infections of humans.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI081690-05
Application #
8586290
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Perdue, Samuel S
Project Start
2010-01-15
Project End
2014-12-31
Budget Start
2014-01-01
Budget End
2014-12-31
Support Year
5
Fiscal Year
2014
Total Cost
$334,556
Indirect Cost
$98,332
Name
University of Minnesota Twin Cities
Department
Zoology
Type
Schools of Earth Sciences/Natur
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Oliver, Jonathan D; Burkhardt, Nicole Y; Felsheim, Roderick F et al. (2014) Motility characteristics are altered for Rickettsia bellii transformed to overexpress a heterologous rickA gene. Appl Environ Microbiol 80:1170-6
Burkhardt, Nicole Y; Baldridge, Gerald D; Williamson, Phillip C et al. (2011) Development of shuttle vectors for transformation of diverse Rickettsia species. PLoS One 6:e29511