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Molecular and Cellular Biology, May 2003, p. 3636-3645, Vol. 23, No. 10
0270-7306/03/$08.00+0     DOI: 10.1128/MCB.23.10.3636-3645.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Nonmuscle Myosin Promotes Cytoplasmic Localization of PBX

He Huang,1,2 Miltiadis Paliouras,3 Isabel Rambaldi,1 Paul Lasko,3 and Mark Featherstone1,2,4,5*

McGill Cancer Centre,1 Division of Experimental Medicine, Department of Medicine,2 Departments of Biology,3 Oncology,4 Biochemistry, McGill University, Montréal, Québec, Canada H3G 1Y65

Received 18 December 2002/ Accepted 11 February 2003

In the absence of MEIS family proteins, two mechanisms are known to restrict the PBX family of homeodomain (HD) transcription factors to the cytoplasm. First, PBX is actively exported from the nucleus via a CRM1-dependent pathway. Second, nuclear localization signals (NLSs) within the PBX HD are masked by intramolecular contacts. In a screen to identify additional proteins directing PBX subcellular localization, we identified a fragment of murine nonmuscle myosin II heavy chain B (NMHCB). The interaction of NMHCB with PBX was verified by coimmunoprecipitation, and immunofluorescence staining revealed colocalization of NMHCB with cytoplasmic PBX in the mouse embryo distal limb bud. The interaction domain in PBX mapped to a conserved PBC-B region harboring a potential coiled-coil structure. In support of the cytoplasmic retention function, the NMHCB fragment competes with MEIS1A to redirect PBX, and the fly PBX homologue EXD, to the cytoplasm of mammalian and insect cells. Interestingly, MEIS1A also localizes to the cytoplasm in the presence of the NMHCB fragment. These activities are largely independent of nuclear export. We show further that the subcellular localization of EXD is deregulated in Drosophila zipper mutants that are depleted of nonmuscle myosin heavy chain. This study reveals a novel and evolutionarily conserved mechanism controlling the subcellular distribution of PBX and EXD proteins.

* Corresponding author. Mailing address: McGill Cancer Centre, McGill University, 3655 Promenade Sir William Osler, Montreal, QC, Canada H3G 1Y6. Phone: (514) 398-8937. Fax: (514) 398-6769. E-mail: mark.featherstone{at}mcgill.ca.

Molecular and Cellular Biology, May 2003, p. 3636-3645, Vol. 23, No. 10
0022-538X/03/$08.00+0     DOI: 10.1128/MCB.23.10.3636-3645.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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