Nonmuscle myosin promotes cytoplasmic localization of PBX.
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.
McGill Cancer Centre, Division of Experimental Medicine, Department of Medicine, McGill University, Montréal, Québec, Canada H3G 1Y6.
He
H
Huang
Miltiadis
M
Paliouras
Isabel
I
Rambaldi
Paul
P
Lasko
Mark
M
Featherstone
D000818:Animals; D019556:COS Cells; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D004330:Drosophila; D005121:Extremities / Q000196:embryology; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008856:Microscopy, Fluorescence; D008957:Models, Genetic; D009218:Myosins / Q000378:metabolism*; D024501:Nonmuscle Myosin Type IIB / Q000378:metabolism; D000075062:Pre-B-Cell Leukemia Transcription Factor 1; D011233:Precipitin Tests; D011401:Promoter Regions, Genetic; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011518:Proto-Oncogene Proteins / Q000737:chemistry* / Q000378:metabolism*; D051381:Rats; D014162:Transfection; D020798:Two-Hybrid System Techniques
D004268:DNA-Binding Proteins; D000075062:Pre-B-Cell Leukemia Transcription Factor 1; D011518:Proto-Oncogene Proteins; C083400:PBX1 protein, human; D024501:Nonmuscle Myosin Type IIB; D009218:Myosins