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Abstract
Membrane morphology is an important structural determinant as it reflects cellular functions. The pentaspan membrane protein Prominin-1 (Prom1/CD133) is known to be localised to protrusions and plays a pivotal role in migration and the determination of cellular morphology; however, the underlying mechanism of its action have been elusive. Here, we performed molecular characterisation of Prom1, focussing primarily on its effects on cell morphology. Overexpression of Prom1 in RPE-1 cells triggers multiple, long, cholesterol-enriched fibres, independently of actin and microtubule polymerisation. A five amino acid stretch located at the carboxyl cytosolic region is essential for fibre formation. The small GTPase Rho and its downstream Rho-associated coiled-coil-containing protein kinase (ROCK) are also essential for this process, and active Rho colocalises with Prom1 at the site of initialisation of fibre formation. In mouse embryonic fibroblast (MEF) cells we show that Prom1 is required for chloride ion efflux induced by calcium ion uptake, and demonstrate that fibre formation is closely associated with chloride efflux activity. Collectively, these findings suggest that Prom1 affects cell morphology and contributes to chloride conductance.
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Details
1 Developmental Biomedical Science, Division of Biological Sciences, Nara Institute of Science and Technology, 8916-5, Takayama-cho, Ikoma, Japan
2 Laboratory for Cell Lineage Modulation, Center for Developmental Biology, Chuo-ku, Kobe, Japan
3 Department of Systems Life Engineering, Maebashi Institute of Technology, Maebashi, Gunma, Japan
4 Faculty of Brain Sciences, UCL Institute of Ophthalmology, University College London, London, United Kingdom
5 Laboratory for Animal Resources and Genetic Engineering, RIKEN Center for Biosystems Dynamics Research, Chuo-ku, Kobe, Japan
6 Department of Ophthalmology, Yamaguchi University School of Medicine, Ube, Japan
7 Laboratory for Cell Lineage Modulation, Center for Developmental Biology, Chuo-ku, Kobe, Japan; Division of Stem Cell Biology, Institute for Genetic Medicine, Hokkaido University, Kita-Ku, Sapporo, Japan