Mammalian knock out cells reveal prominent roles for atlastin GTPases in ER network morphology
| Autor: | Peng-Peng Zhu, Benoît Renvoisé, Seonghee Park, Lymarie Maldonado-Báez, Craig Blackstone, Guohua Zhao |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Atlastin Hereditary spastic paraplegia Cell Survival Biology Bone morphogenetic protein Endoplasmic Reticulum GTP Phosphohydrolases 03 medical and health sciences symbols.namesake Gene Knockout Techniques Mice 0302 clinical medicine Viral Envelope Proteins medicine Adipocytes Animals Humans Gene knockout Cell Proliferation Mammals Membrane Glycoproteins Endoplasmic reticulum Cell Cycle Cell Differentiation Cell Biology Golgi apparatus medicine.disease Phenotype Cell biology 030104 developmental biology HEK293 Cells Bone Morphogenetic Proteins symbols Unfolded protein response NIH 3T3 Cells 030217 neurology & neurosurgery Signal Transduction |
| Zdroj: | Experimental cell research. 349(1) |
| ISSN: | 1090-2422 |
| Popis: | Atlastins are large, membrane-bound GTPases that participate in the fusion of endoplasmic reticulum (ER) tubules to generate the polygonal ER network in eukaryotes. They also regulate lipid droplet size and inhibit bone morphogenetic protein (BMP) signaling, though mechanisms remain unclear. Humans have three atlastins (ATL1, ATL2, and ATL3), and ATL1 and ATL3 are mutated in autosomal dominant hereditary spastic paraplegia and hereditary sensory neuropathies. Cellular investigations of atlastin orthologs in most yeast, plants, flies and worms are facilitated by the presence of a single or predominant isoform, but loss-of-function studies in mammalian cells are complicated by multiple, broadly-expressed paralogs. We have generated mouse NIH-3T3 cells lacking all three mammalian atlastins (Atl1/2/3) using CRISPR/Cas9-mediated gene knockout (KO). ER morphology is markedly disrupted in these triple KO cells, with prominent impairment in formation of three-way ER tubule junctions. This phenotype can be rescued by expression of distant orthologs from Saccharomyces cerevisiae (Sey1p) and Arabidopsis (ROOT HAIR DEFECTIVE3) as well as any one of the three human atlastins. Minimal, if any, changes are observed in the morphology of mitochondria and the Golgi apparatus. Alterations in BMP signaling and increased sensitivity to ER stress are also noted, though effects appear more modest. Finally, atlastins appear required for the proper differentiation of NIH-3T3 cells into an adipocyte-like phenotype. These findings have important implications for the pathogenesis of hereditary spastic paraplegias and sensory neuropathies associated with atlastin mutations. |
| Databáze: | OpenAIRE |
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