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Science 306 (5700): 1370-1373

Copyright © 2004 by the American Association for the Advancement of Science

Regulated Fast Nucleocytoplasmic Shuttling Observed by Reversible Protein Highlighting

Ryoko Ando, Hideaki Mizuno, Atsushi Miyawaki*

Abstract: The observation of the regulation of fast protein dynamics in a cellular context requires the development of reliable technologies. Here, a signal regulation cascade reliant on the stimulus-dependent acceleration of the bidirectional flow of mitogen-activated protein kinase (extracellular signal-regulated kinase) across the nuclear envelope was visualized by reversible protein highlighting. Light-induced conversion between the bright and dark states of a monomeric fluorescent protein engineered from a novel coral protein was employed. Because of its photochromic properties, the protein could be highlighted, erased, and highlighted again in a nondestructive manner, allowing direct observation of regulated fast nucleocytoplasmic shuttling of key signaling molecules.

Laboratory for Cell Function and Dynamics, Advanced Technology Development Group, Brain Science Institute, RIKEN, 2-1 Hirosawa, Wako-city, Saitama, 351-0198, Japan.

* To whom correspondence should be addressed. E-mail: matsushi{at}brain.riken.jp


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