Rockefeller University
To study how mechanical force impacts the fidelity of transcription and coordinates the development of multicellular structures
Unraveling how mechanical forces / physical effects contribute to biological function is an underexplored yet important aspect of understanding living systems. This grant provides continuing support to a trio of early-career researchers at Rockefeller University for a series of experiments geared at understanding how mechanical force impacts two important biological functions: the copying of information stored in DNA (transcription) and the coordinated development of a field of cells into a multicellular functional unit (here, skeletal tissue). The proposed transcription research will study how force impacts the dynamics and fidelity of the primary biomolecular machine responsible for transcription, RNA polymerase (RNAP). Forces will be applied to RNAP either using laser tweezers or via collisions between RNAP and various biomolecules that mimic RNAP collisions in live cells. Fluorescence microscopy will capture RNAP dynamics and cryogenic electron microscopy (cryo-EM) will reveal how the molecular-scale structure, and therefore the biochemical activity, of RNAP is modified by collisions. The tissue development research will use cryo-EM along with cellular biology methods that stimulate cell contraction -and thus force propagation- to study changes in molecular architecture that are driven by supracellular (beyond a single cell) force transmission.