Streptococcus pneumoniae (pneumococcus) is an important human pathogen and the most common cause of bacterial meningitis, pneumonia and otitis media. Although it can cause significant disease, pneumococcus is part of the normal nasopharyngeal flora. Its ability to take up and incorporate foreign DNA has allowed pneumococcus to resist attempts at clearance by providing a resource for genetic adaptation. Genome analysis has demonstrated that this species is characterized by an enormous amount of genetic diversity largely derived from recombination with DNA from other streptococci. The competence state in pneumococcus is regulated by a quorum sensing system controlled by the com system which drives the production of a large number of genes including those involved in DNA uptake and recombination. Competent pneumococci can lyse non-competent members of the population releasing their DNA via a mechanism called fratricide. Released DNA is taken up by competent cells and can serve as a source of new genetic material. The proteins involved in fratricide and fratricide immunity are highly conserved among pneumococci and other related strep species limiting the target population for lysis to members of the population that have failed to induce competence. The production of lytic pneumococcal bacteriocins (pneumocins) from the blp locus has the potential to expand the target range for DNA release. The blp locus is controlled by a parallel quorum sensing system to that required for competence induction. A number of genes in the blp locus have been shown to be upregulated during early competence induction and we have demonstrated that stimulation of competence induces transcription of the pneumocin genes in some isolates. Because the pneumocins and their immunity proteins encoded in the blp cluster are variable from strain to strain, they can promote lysis of a wide range of pneumococci and related streptococcal species independent of the competence state of the target. We hypothesize that coordinate expression of pneumocins with the competence state will increase the potential target organisms for DNA release and uptake.
In Aim1 of the proposal we will investigate the mechanism of cross stimulation of pneumocin by competence induction.
In Aim 2 we will determine the role of the regulatory proteins CiaRH and HtrA in controlling the cross talk between the blp and com systems.
In Aim3, we will verify that coordinate blp/com regulation increases the target range for DNA release by demonstrating that pneumocin producing strains have an advantage over strains that rely solely on fratricide in the acquisition of foreign DNA. DNA exchange events will be studied in vitro under optimized conditions and in vivo, during mouse nasal colonization. The persistence of this pathogen despite ongoing attempts at clearance from both the host and the surrounding flora is a testament to the power of genomic diversity and adaptation. Understanding how the variably expressed pneumocins contribute to this diversity will allow us to better anticipate and perhaps block attempts at escape from control measures via genetic exchange.

Public Health Relevance

Streptococcus pneumoniae has persisted as a major human pathogen despite targeted attempts at eradication through vaccination and antibiotic use. This is largely due to its ability to alter its genetic content via DNA exchange. Genetic adaptation requires acquisition of new pieces of DNA that may provide an advantage to the bacterium in the form of antibiotic resistance or changes in surface structures that are recognized by antibodies. This project involves understanding the role that bacteriolytic peptides called pneumocins play in increasing genetic diversity and adaptation.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI101285-02
Application #
8613433
Study Section
Bacterial Pathogenesis Study Section (BACP)
Program Officer
Taylor, Christopher E,
Project Start
2013-02-05
Project End
2018-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
2
Fiscal Year
2014
Total Cost
$344,468
Indirect Cost
$119,468
Name
University of Michigan Ann Arbor
Department
Pediatrics
Type
Schools of Medicine
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109