Molecular function of Myosin-l

University Of Pennsylvania, Philadelphia PA

Investigators

Linked publications & trials

Abstract

SUMMARY The goal of the parent R37 project is to determine the molecular mechanisms of the myosin-I family of motors. Myosin-Is comprise the largest unconventional myosin family found in humans, and its large size and expression profile distinguish it as one of the most diverse. Myosin-Is physically link cell membranes to the underlying actin cytoskeleton where they play essential roles in powering membrane dynamics, membrane trafficking, and mechanical signal-transduction. Discoveries made during the tenure of this award include revealing (a) synergies between myosin- I and Arp2/3 mediated actin polymerization, and (b) the mechanisms by which non-muscle tropomyosins regulate myosin-I activity and subcellular localization. Exciting new experiments will provide the mechanistic details necessary to understand how these proteins use the mechanochemical cycle of myosin to bring about changes in membrane structure and dynamics. To accelerate progress of our Aims, we require the ability to quantify the binding, kinetics, and distribution of fluorescently labeled proteins at the single-molecule level via TIRF microscopy. Our current equipment is built on a > 20-year-old microscope that does not have the ability to acquire data with the positional and temporal resolution or wavelengths necessary to address our questions. Additionally, the software running the system is no longer supported by the manufacturer, which will leave us without crucial technical support, making the system obsolete. The requested TIRM microscope will allow us to rapidly achieve our Aims, and we anticipate that the instrument will be used daily by multiple investigators in the lab over a several year period, coincident with the funding of the R37.