Publisher Summary Nuclear shuttling is used to regulate the onset of cell cycle events and the access of cytoplasmic proteins to the cell periphery. Some proteins may shuttle continuously in and out of the nucleus, whereas others may undergo regulated entry and exit. The existence of nuclear shuttling as a regulatory mechanism for a particular protein may not always be obvious from the localization pattern of the protein at steady state. This chapter describes a variety of ways in which it can be evaluated whether a protein that appears static at steady state actually shuttles through the nucleus. Demonstrating that a nuclear protein shuttles involves finding conditions that reveal its ability to be exported out of the nucleus. A protein that appears to be cytoplasmic at steady state may also shuttle, but the rate of nuclear export may be greater than the rate of nuclear import. Demonstrating that a cytoplasmic protein shuttles involves finding conditions that clearly reveal the existence of a nuclear pool that is exported.
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