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doi: 10.1242/10.1242/jcs.00095

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Journal of Cell Science 115, 4167-4176 (2002)
Copyright © 2002 The Company of Biologists Limited
doi: 10.1242/jcs.00095

Research Article

Transcription factors RUNX1/AML1 and RUNX2/Cbfa1 dynamically associate with stationary subnuclear domains

Kimberly S. Harrington1, Amjad Javed1, Hicham Drissi1, Sandra McNeil1, Jane B. Lian1, Janet L. Stein1, André J. van Wijnen1, Yu-Li Wang1,2 and Gary S. Stein1,*

1 Department of Cell Biology and Cancer Center, University of Massachusetts Medical School, Worcester, Massachusetts 01655-0106, USA
2 Department of Physiology, University of Massachusetts Medical School, Worcester, Massachusetts 01655-0106, USA

* Author for correspondence (e-mail: gary.stein{at}umassmed.edu)

Accepted 5 August 2002

The runt-related transcription factors (RUNX/Cbfa/AML) are essential for cellular differentiation and fetal development. C-terminal truncations of RUNX factors that eliminate the targeting of these factors to subnuclear foci result in lethal hematopoietic and skeletal phenotypes. Here we demonstrate that in living cells the RUNX C-terminus is necessary for the dynamic association of RUNX into stable subnuclear domains. Time-lapse fluorescence microscopy shows that RUNX1 and RUNX2 localize to punctate foci that remain stationary in the nuclear space. By fluorescence recovery after photobleaching assays, both proteins are shown to dynamically associate at these subnuclear foci, with a 10 second half-time of recovery. A truncation of RUNX2, removing its intranuclear targeting signal (NMTS), increases its mobility by an order of magnitude, resulting in a half-time of recovery equivalent to that of EGFP alone. We propose that the dynamic shuttling of RUNX factors in living cells to positionally stabilized foci, which is dependent on the C-terminus, is a component of the mechanism for gene regulation in vivo.

Key words: Intranuclear targeting, Runt homology factors, Green fluorescent protein, Fluorescence recovery after photobleaching, Nuclear matrix




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© The Company of Biologists Ltd 2002