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Michael Sheetz, James Spudich, and Ronald Vale Receive the 2012 Albert Lasker Basic Medical Research Award

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Michael Sheetz, James Spudich, and Ronald Vale Receive the 2012 Albert Lasker Basic Medical Research Award Molecules in motion: Michael Sheetz, James Spudich, and Ronald Vale receive the 2012 Albert Lasker Basic Medical Research Award Sarah Jackson J Clin Invest. 2012;122(10):3374-3377. https://doi.org/10.1172/JCI66361. News The Albert and Mary Lasker Foundation honors three pioneers in the field of molecular motor proteins, Michael Sheetz, James Spudich, and Ronald Vale (Figure 1), for their contribution to uncovering how these proteins catalyze movement. Sheetz and Spudich developed the first biochemical assay to reconstitute myosin motor activity and showed that myosin and ATP alone were sufficient to direct transport along actin filaments. Fueled by this discovery, Vale and Sheetz examined transport in giant squid axon extracts and discovered a new motor, kinesin, that moves along microtubules. Cumulatively, their work opened the door for understanding how motor proteins drive transport of a host of molecules involved in numerous cellular processes, including cell polarity, cell division, cellular movement, and signal transduction. Muscle movement Much of the early research on cellular movement focused on understanding muscle contraction. Filaments of actin and myosin form a banding pattern in muscle that is readily visible by light microscopy, with a characteristic striated appearance. The functional unit of muscle, termed a sarcomere, repeats at regular intervals along the muscle fibers. Measurements from contracted, resting, and stretched muscle using high-resolution light microscopy supported the notion that contraction was due to the sliding of actin and myosin filaments (1, 2). In 1969, preeminent British scientist Hugh Huxley proposed that crossbridges that connect actin and myosin generate the sliding […] Find the latest version: https://jci.me/66361/pdf News Molecules in motion: Michael Sheetz, James Spudich, and Ronald Vale receive the 2012 Albert Lasker Basic Medical Research Award The Albert and Mary Lasker Founda- James Spudich joined the Huxley labora- motility along actin. He and his colleagues tion honors three pioneers in the field of tory as a young postdoctoral researcher. took a multipronged approach, using molecular motor proteins, Michael Sheetz, From his experiences there, Spudich was biophysical, biochemical, and genetic James Spudich, and Ronald Vale (Figure 1), keenly aware of the need for a biochemical techniques to uncover the mechanism of for their contribution to uncovering how assay for the function of interest, move- myosin action. Along with postdoctoral these proteins catalyze movement. Sheetz ment, and to explore the mechanism of fellow Margaret Clarke, Spudich identi- and Spudich developed the first biochem- energy transduction by the myosin pro- fied a model organism for studying non- ical assay to reconstitute myosin motor tein. In a recent interview with the JCI, muscle actin and myosin biochemistry, activity and showed that myosin and ATP Spudich recalled that many of the bio- the amoeba Dictyostelium (4). Early studies alone were sufficient to direct transport physical experiments from the 1970s did with the organism involved actin filaments along actin filaments. Fueled by this dis- not support the swinging crossbridge attached to tiny polystyrene beads, which covery, Vale and Sheetz examined trans- model, “Frankly, people were beginning to provided encouragement for an in vitro port in giant squid axon extracts and dis- think that it may not work the way Hugh motility assay. Years later, the Spudich covered a new motor, kinesin, that moves had proposed.” When Spudich branched laboratory remarkably developed genetic along microtubules. Cumulatively, their off and opened his own research labora- tools to examine Dictyostelium cells lacking work opened the door for understanding tory, he set out to rigorously and defini- myosin and to test the ability of various how motor proteins drive transport of a tively test models of energy transduction myosin mutants to compensate. Spudich host of molecules involved in numerous by myosin. In addition, he developed a recalled, “My graduate student Arturo cellular processes, including cell polarity, strong interest in the nascent field of non- De Lozanne unexpectedly discovered that cell division, cellular movement, and sig- muscle myosins. one could use the genetic approach of nal transduction. homologous recombination with Dictyo- Recapitulating motion stelium. That did a lot for us because we Muscle movement in the laboratory were then able to create a cell that did not Much of the early research on cellular First at the University of California, San contain myosin. That cell was unable to movement focused on understanding Francisco, and later at Stanford, a pri- divide into two daughter cells (5). Then, muscle contraction. Filaments of actin mary goal in Spudich’s laboratory was we made mutations in the myosin and and myosin form a banding pattern in to develop an in vitro assay using only put those mutated myosins back into the muscle that is readily visible by light purified components to examine myosin cell that lacked the myosin to follow the microscopy, with a characteristic striated appearance. The functional unit of mus- cle, termed a sarcomere, repeats at regular intervals along the muscle fibers. Mea- surements from contracted, resting, and stretched muscle using high-resolution light microscopy supported the notion that contraction was due to the sliding of actin and myosin filaments (1, 2). In 1969, preeminent British scientist Hugh Huxley proposed that crossbridges that connect actin and myosin generate the sliding motion that moves actin during muscle contraction (3). His model, termed the swinging crossbridge hypothesis, pre- dicted that these crossbridges bind and release in a cyclical manner to produce the force that causes actin sliding, thus propelling movement and muscle con- traction. While an elegant model, it was difficult to test and remained controver- Figure 1 sial for many years. Soon after the swing- Ronald Vale (left), Michael Sheetz (center), and James Spudich (right) are the winners of the ing crossbridge hypothesis was proposed, 2012 Albert Lasker Basic Medical Research Award. 3374 The Journal of Clinical Investigation http://www.jci.org Volume 122 Number 10 October 2012 news supported by actin cables. We could very vitro. The images and movies they had readily monitor movement in this system looked like, perhaps, how axonal trans- by light microscopy, so it seemed like an port might work. The three of us began ideal system.” The idea was a success, and talking and that seemed like an exciting their work showed for the first time clear possibility.” In addition to the work of movement of myosin along actin filaments Sheetz and Spudich on myosin, reports in an ATP-dependent manner. by Robert Allen and Scott Brady demon- Spudich’s laboratory was still focused, strated abundant transport of organ- however, on developing an in vitro system elles and vesicles within axons, using Figure 2 using purified actin rather than relying on an approach that combined video with An in vitro assay to test myosin motility. the complex Nitella-derived actin substra- microscopy to monitor giant squid axons Michael Sheetz and James Spudich devel- tum. They returned to earlier attempts to (8, 9). Building upon these studies, Vale oped the first biochemical assay to examine reconstitute a totally defined system. In and Sheetz began a serious collaboration the movement of myosin along actin filaments. this approach, now involving graduate stu- to understand the molecular motors driv- Using myosin bound to fluorescently labeled dent Stephen Kron, actin filaments were ing motility in squid axons. beads, they tracked myosin movement micro- scopically along actin-aligned filaments, first conjugated to biotin and fixed on avidin- Sheetz and Vale went to the Marine Bio- in the alga Nitella (6) and later using purified coated slides, with buffer flow orienting logical Laboratory at Woods Hole, where actin bound to glass slides (7). Spudich and the actin filaments (Figure 3). Finally, they they had ready access to giant squid. Using colleagues later showed that the S1 head of were able to recreate myosin movement a cellular extrusion from axons known myosin both binds to actin and hydrolyzes entirely in the test tube, using pure actin, as axoplasm, they teamed up with elec- ATP to generate motion (14). pure myosin, and ATP (7). tron microscopists, Bruce Schnapp and Thomas Reese, to monitor motility. Ini- A new way of moving tially, Vale and Sheetz thought that non- behavior in vivo that resulted from those Just a few floors away from Spudich and muscle myosins might also drive motion different mutations.” Despite much prog- Sheetz in the Fairchild Science Building in axons. However, they were surprised ress, several technical hurdles left the Spu- at Stanford, Ronald Vale was studying the to find that the molecular motor driving dich laboratory still searching for an assay development of the nervous system and motion in squid axoplasm appeared quite to reconstitute myosin movement along nerve regeneration. Knowing that axons different from myosin. When Sheetz and actin with known, purified components. can reach up to a meter in length, Vale was Vale tested whether myosin conjugated A major breakthrough in the develop- deeply interested in understanding how to beads could move in squid axoplasm, ment of an in vitro assay for myosin move- long-distance transport occurs between they saw no movement. “Since the initial ment along actin occurred when Michael the cell body and terminus of nerve cells. idea wasn’t working, we decided to take Sheetz joined the Spudich laboratory in Vale recognized that the work of Spudich a more naive approach and really dissect 1982. Spudich recalled, “Our model was and Sheetz had important implications the system step by step to figure out what that actin and myosin would move with- for other kinds of cellular transport. In elements of the cytoskeleton were work- out anything else, but there was no proof an interview with the JCI, Vale recalled, “I ing. Maybe it wasn’t actin, and indeed it that that was the way nature worked.
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