School of Dentistry
Twin Cities
These researchers study the mechanism of force generation of ring ATPase motors using the bacteriophage phi29 as a model system. During dsDNA virus assembly, DNA is packaged into a preformed capsid using powerful ring ATPases, which are members of a broadly distributed class of polymer translocating motors found in all living systems, the ASCE family. Several decades of high-resolution structure determination, single-molecule biophysical studies, long-timescale simulation, and traditional genetics and biochemistry have converged, making the phi29 DNA packaging motor one of the best described of these systems. Using a CRISPR-based approach, this group has begun to produce libraries of phages that contain randomized mutations at amino acid positions of interest in the ATPase; subsequent infection of these libraries yields only phages with amino acid substitutions that can be tolerated, thus allowing for the rapid screening of mutations for their viability. Next-gen sequencing will allow the researchers to sequence the population of phages produced during these experiments; they use MSI for data acquisition, delivery, and analysis.