The research will first focus on what are known as master cytoskeletal regulatory proteins (CRPs) that control how cells take shape and move. The overexpression of these proteins is known to be associated with poor prognosis for some cancer patients, but the researchers said their workings remain largely unknown.
Balzsi's lab has developed the ability to prompt the expression of genetically modified proteins that give the researchers a level of control over CRPs with minimal disruption of other cell functions. This lets the researchers design experiments to see how the systems inside cells respond to stress.
Diehl is most interested in a CRP called "IQGAP." "It both tunes cell-signaling responses and is involved in controlling proteins that regulate the cytoskeleton," he said. "It's at the junction of those two behaviors." By perturbing the regulatory network through modified IQGAP, "we can look at the network response and correlate that with cellular behavior. It's a very powerful approach that can be applied to many molecular and cellular processes."
Diehl's lab will view those processes with a multiplexed "super resolution" imaging technique that lets researchers see individual protein molecules inside cells in three dimensions. That will allow them to identify multiple proteins in a cell, perhaps hundreds, at once. Key to the technique is the ability to combine images that focus on different target proteins into a comprehensive picture of cellular structure.
"We have developed erasable molecular imaging probes for these types of studies," Diehl said. Tagging proteins with fluorescent dyes allows researchers to "see" them, but current microscopes are limited to only a few color channels. With the ability to tag, erase and retag proteins, the Diehl lab can capture much more information from snapshots of a cell taken before and after perturbation.
Qutub's lab analyze
|Contact: David Ruth|