Intellectual Merit. Conjugation involves the transfer of DNA directly from a donor cell to a recipient cell. Since conjugative elements can deliver their own genetic material as well as other resident DNA, conjugation has played a profound role in bacterial evolution, allowing organisms to acquire new genes and adapt to new environmental niches. During conjugation, DNA is transferred from cell to cell through a specialized DNA translocation channel that is encoded by the conjugative element. This project will characterize the conserved ATPase ConE, a critical energetics component of the DNA translocation channel of the conjugative element ICEBs1 of Bacillus subtilis. This project is designed to reveal which conjugation proteins interact with and recruit ConE to the cell membrane. Furthermore, the sequences of ConE required for conjugation, ATP hydrolysis, and localization to the membrane will be explored. These studies should lead to the discovery of other critical components of the ICEBs1 conjugation machinery and a better understanding of the fundamental principles that govern horizontal gene transfer in bacteria.
Broader Impacts. Major impacts of this project are to enhance undergraduate student training while broadening our understanding of horizontal gene transfer. The research will be designed, conducted, and analyzed by undergraduate students with guidance from the PI. The PI will continue to integrate inquiry-based research into her undergraduate biochemistry laboratory course and high school outreach activities. Furthermore, this project will permit the purchase of a research-grade fluorescence microscope which will allow students to follow a wider range of research interests and learn sophisticated microscopy techniques. The percentage of students entering top-tier graduate programs and careers in the biosciences is expected to increase, as many more students will have the opportunity to gain useful research skills, present their work, and experience the excitement of scientific discovery.
