In bacteria, genetic information is transferred "vertically" because DNA is inherited by daughter cells. Genetic information can also be transferred "horizontally", accounting in many cases for the exchange of genes that contribute to virulence and antibiotic resistance. Horizontal transfer is a major factor in the evolution of bacteria. An important route of horizontal transfer is by the release of DNA into the environment and the subsequent uptake of this DNA by bacteria, a process known as transformation. The work proposed in this application is devoted to obtaining a mechanistic understanding of the transformation process. We wish to learn how cells bind DNA to their surface and how they internalize this DNA, the role of a filamentous structure known as the pseudopilus in the uptake process and to use microscopy to visualize the uptake and processing of the DNA once it is internalized as well as the interaction of the DNA with the recipient cell chromosome. Cells that are capable of being transformed are in a physiological state in which their growth, division and DNA replication are halted. We will continue our efforts to understand the causes of this arrest and the mechanisms that enable the cells to eventually resume growth.

Public Health Relevance

Horizontal gene transfer is an important route for the spread of antibiotic resistance and virulence genes. An important mechanism of horizontal transfer is transformation, the uptake of DNA previously released into the environment. We wish to gain a mechanistic understanding of the transformation process.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM043756-24
Application #
8646922
Study Section
Prokaryotic Cell and Molecular Biology Study Section (PCMB)
Program Officer
Deatherage, James F
Project Start
1990-12-01
Project End
2017-02-28
Budget Start
2014-03-01
Budget End
2015-02-28
Support Year
24
Fiscal Year
2014
Total Cost
$397,500
Indirect Cost
$147,500
Name
Rutgers University
Department
Public Health & Prev Medicine
Type
Schools of Medicine
DUNS #
078795851
City
Newark
State
NJ
Country
United States
Zip Code
07103
Briley Jr, Kenneth; Dorsey-Oresto, Angella; Prepiak, Peter et al. (2011) The secretion ATPase ComGA is required for the binding and transport of transforming DNA. Mol Microbiol 81:818-30
Briley Jr, Kenneth; Prepiak, Peter; Dias, Miguel J et al. (2011) Maf acts downstream of ComGA to arrest cell division in competent cells of B. subtilis. Mol Microbiol 81:23-39
Burton, Briana; Dubnau, David (2010) Membrane-associated DNA transport machines. Cold Spring Harb Perspect Biol 2:a000406
Dubnau, David (2010) Swim or chill: lifestyles of a bacillus. Genes Dev 24:735-7
Johnsen, P J; Dubnau, D; Levin, B R (2009) Episodic selection and the maintenance of competence and natural transformation in Bacillus subtilis. Genetics 181:1521-33
Kramer, Naomi; Hahn, Jeanette; Dubnau, David (2007) Multiple interactions among the competence proteins of Bacillus subtilis. Mol Microbiol 65:454-64
Chen, Ines; Provvedi, Roberta; Dubnau, David (2006) A macromolecular complex formed by a pilin-like protein in competent Bacillus subtilis. J Biol Chem 281:21720-7
Draskovic, Irena; Dubnau, David (2005) Biogenesis of a putative channel protein, ComEC, required for DNA uptake: membrane topology, oligomerization and formation of disulphide bonds. Mol Microbiol 55:881-96
Chen, Ines; Christie, Peter J; Dubnau, David (2005) The ins and outs of DNA transfer in bacteria. Science 310:1456-60
Hahn, Jeanette; Maier, Berenike; Haijema, Bert Jan et al. (2005) Transformation proteins and DNA uptake localize to the cell poles in Bacillus subtilis. Cell 122:59-71

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