@article{THESIS,
      recid = {1772},
      author = {Krenc, Agata Karolina},
      title = {FLUORESCENCE INTERFERENCE CONTRAST MICROSCOPY, A NEW TOOL  TO STUDY COLLECTIVE MOTOR DYNAMICS},
      publisher = {University of Chicago},
      school = {Ph.D.},
      address = {2016-08},
      number = {THESIS},
      pages = {98},
      abstract = {Teams of molecular motors called myosins carry out  intracellular transport and contract the actin  cytoskeleton. To fully understand the behavior of  multi-myosin ensembles we need to know the properties of  individual myosins and the mode of interaction between  them. Current models of the interactions within the myosin  complex treat the actin filament as a stiff rod, not  contributing to the regulation of collective myosin  dynamics. Here, I present data suggesting that force  transduction through the actin filament is an important  element of interaction within myosin-6 ensembles in vitro.  Multiple myosin-6s coordinate their steps if they are  separated by a short (and therefore high-force bearing)  segment of actin. The measurements were performed using  Fluorescence Interference Contrast Microscopy (FLIC). In  FLIC, the fluorescence intensity is proportional to the  height of a fluorophore above a reflective surface. FLIC  allows for the detection of small changes in a height of  fluorescently labeled actin.  Using FLIC, I was able to  assign the positions of myosins in a gliding filament assay  geometry and to measure the time each of them spends bound  to actin (the attachment time). I show that myosin-6 holds  actin about 10 nm above the surface. However, due to  asynchronous myosin stepping, frequent buckles up to about  70 nm high appear. The buckle lifetime decreases as the  distance between the myosin-6s is reduced, a sign of  inter-motor coordination. Similar measurements were  performed for myosin-5 and NMIIB, showing that the FLIC  assay is a robust and versatile technique allowing for  ensemble measurements with single molecule insight.},
      url = {http://knowledge.uchicago.edu/record/1772},
      doi = {https://doi.org/10.6082/uchicago.1772},
}