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Rockefeller researchers discover a biological clock within a clock

at the two proteins idle in a cell's cytoplasm until they bump into each other and then, bound together, enter the nucleus. But Young and Pablo Meyer, who was then a graduate student in Young's lab, used a novel method to show that this scenario was far too simple.

Meyer, a physicist by training, found himself frustrated by how little he could see of what was occurring in a cell. "The truth is, we really don't know, mechanistically, what happens in the cytoplasm, and how things are being done in such a precise way," Meyer says. So he turned to a technique invented in 1948, called fluorescence resonance energy transfer; FRET gauges interactions between proteins by fluorescently tagging them and measuring how they react to different wavelengths of light. But although the technique can provide useful information, it's so complicated that researchers rarely use it. And no one had ever thought to use it to follow proteins in a single cell for an extended period of time.

"This begins to measure all these biochemical interactions inside the cell," Meyer says. For the first time, Period and Timeless could be tracked within a cell for eight hours or longer. "No one had ever labeled the components to follow them over time, to see one clock as it ticks away in a single cell," Young says. "All the biochemistry and molecular biology that had been done on this had been piecing together information from dead flies." But now, instead of freeze-frames, they had a movie.

The movie allowed them to follow the interactions between Period and Timeless with a resolution never before possible. They discovered that, rather than randomly colliding, the two proteins bind together in the cytoplasm almost immediately and create what Young and Meyer refer to as an "interval timer." Then, six hours after coming together, the complexes rapidly break apart and the proteins move into the nucleus singly, all of them within minutes of each other. "Some switch is thrown at six
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Contact: Joseph Bonner
bonnerj@rockefeller.edu
212-327-8998
Rockefeller University
12-Jan-2006


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