DNA-Mediated Assembly of Functional Chemoreceptor Complexes in E. coli

Presenter

Sarah Tobia

Campus

UMass Amherst

Sponsor

Lynmarie K. Thompson, Department of Chemistry, UMass Amherst

Schedule

Session 4, 2:30 PM - 3:15 PM [Schedule by Time][Poster Grid for Time/Location]

Location

Poster Board A70, Campus Center Auditorium, Row 4 (A61-A80) [Poster Location Map]

Abstract

Chemotaxis is the process by which an organism moves in response to a chemical stimulus. The chemotaxis signaling pathway in E. coli relies on a complex array of transmembrane chemoreceptors with cytoplasmic binding partners to transmit a signal and produce a response. The specific mechanisms and dynamics of signaling are not fully understood, and experimentation is limited by the ability to reconstitute the array’s quaternary structure in vitro. Current assembly methods of these complexes achieve homogenous native assembly and allow for activity; however, they lack the precise control necessary to determine how the array and receptors control kinase activity and signal transmission. Using a bi-tetrahedron DNA-nanotechnology scaffold, precise assembly of complexes can be achieved. DNA oligonucleotides extended from the tetrahedron are covalently linked to an N-terminal, cys-modified receptor protein. The DNA tetrahedron with oligonucleotide extensions has been functionally assembled, and successful linkage between the oligonucleotide extensions and the protein has been achieved. Optimizing this product formation will allow for the formation of functional chemoreceptor complexes with the ability to direct assembly and control the architecture of the array. While these techniques are currently being tested for in vitro assembly to study structure and function, modifications can be made to the system to allow for in vivo applications of sensing and delivery.

Keywords

Chemotaxis, Assembly methods, DNA-nanotechnology, Protein interactions and signaling

Research Area

Chemistry and Materials Science

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