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1 Department of Internal Medicine, Washington University School of Medicine, Saint Louis, MO, USA
2 Department of Biochemistry and Molecular Biology and Institute for Genetic Medicine, Keck School of Medicine, University of Southern California, Los Angeles, CA, USA
3 Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA, USA
4 Department of Medicine and Will Rogers Institute of Pulmonary Research Center, University of Southern California, Los Angeles, CA, USA
* To whom correspondence should be addressed. E-mail: brodys{at}msnotes.wustl.edu.
Factors required for commitment of an undifferentiated airway epithelial cell to a ciliated cell are unknown. Cell ultrastructure analysis indicates ciliated cell commitment activates a multi-stage program involving synthesis of cilia precursor proteins and assembly of macromolecular complexes. Foxj1 is an f-box transcription factor expressed in ciliated cells and shown to be required for cilia formation by gene deletion in a mouse model. To identify a specific role for foxj1 in directing the ciliated cell phenotype, we evaluated the capacity of foxj1 to induce ciliogenesis and direct cilia assembly. In a primary culture model of wild type mouse airway epithelial cells, foxj1 expression preceded the appearance of cilia and in cultured foxj1 null cells cilia did not develop. Delivery of foxj1 to polarized epithelial cell lines and primary cultured alveolar epithelial cells failed to promote ciliogenesis. Similarly, delivery of foxj1 to wild type airway epithelial cells did not enhance the total number of ciliated cells. In contrast, delivery of foxj1 to null cells resulted in the appearance of cilia. Analysis revealed that in the absence of foxj1, null cells contained cilia precursor basal bodies, indicating prior commitment to ciliogenesis. However, the basal bodies were disorganized within the apical compartment and failed to dock with the apical membrane. Reconstitution of foxj1 in null cells restored normal basal body organization resulting in axoneme growth. Thus, foxj1 functions in late stage ciliogenesis to regulate programs promoting basal body docking and axoneme formation in cells previously committed to the ciliated cell phenotype.
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