Protein architecture and emergent mechanisms of kinetochore force generation
Our lab studies the biophysical mechanisms of force generation and signaling by the kinetochore, a macromolecular machine that is responsible for genome inheritance. Two fundamental questions about the molecular mechanisms underlying kinetochore function remain unanswered:
(1) How does it generate microtubule polymerization-coupled force?
(2) How does it transduce the mechanical event of microtubule attachment into a diffusible signal?
Answers to these questions have so far eluded investigations mainly because of the macromolecular nature of the kinetochore. The kinetochore relies on emergent properties of at least eight different protein complexes that act in a nanoscale architecture and show cooperative function both in space and time.
Our lab specializes in using genetically encoded fluorescent probes to measure molecular properties in live cell. Our tool-kit includes quantitative microscopy, super-resolution microscopy, FRET, fluorescence polarization microscopy. We are also using spectroscopic methods such as FCS, FCCS, and FLIM in vivo as well as in vitro using recombinant proteins. This technical toolkit allows us to measure molecular properties in vivo. Combined with molecular biology and genetics, we are in a unique position to conduct a systematic “architecture-function” analysis. By defining not only molecular properties but also their nanoscale architecture, we will answer the fundamental questions posed above that lie at the interface between cell biology and biophysics.