RRC ID 54940
Author Mizotani Y, Suzuki M, Hotta K, Watanabe H, Shiba K, Inaba K, Tashiro E, Oka K, Imoto M.
Title 14-3-3εa directs the pulsatile transport of basal factors toward the apical domain for lumen growth in tubulogenesis.
Journal Proc. Natl. Acad. Sci. U.S.A.
Abstract notochord has emerged as a simple and tractable in vivo model for tubulogenesis. Here, using a chemical genetics approach, we identified UTKO1 as a selective small molecule inhibitor of notochord tubulogenesis. We identified 14-3-3εa protein as a direct binding partner of UTKO1 and showed that 14-3-3εa knockdown leads to failure of notochord tubulogenesis. We found that UTKO1 prevents 14-3-3εa from interacting with ezrin/radixin/moesin (ERM), which is required for notochord tubulogenesis, suggesting that interactions between 14-3-3εa and ERM play a key role in regulating the early steps of tubulogenesis. Using live imaging, we found that, as lumens begin to open between neighboring cells, 14-3-3εa and ERM are highly colocalized at the basal cortex where they undergo cycles of accumulation and disappearance. Interestingly, the disappearance of 14-3-3εa and ERM during each cycle is tightly correlated with a transient flow of 14-3-3εa, ERM, myosin II, and other cytoplasmic elements from the basal surface toward the lumen-facing apical domain, which is often accompanied by visible changes in lumen architecture. Both pulsatile flow and lumen formation are abolished in larvae treated with UTKO1, in larvae depleted of either 14-3-3εa or ERM, or in larvae expressing a truncated form of 14-3-3εa that lacks the ability to interact with ERM. These results suggest that 14-3-3εa and ERM interact at the basal cortex to direct pulsatile basal accumulation and basal-apical transport of factors that are essential for lumen formation. We propose that similar mechanisms may underlie or may contribute to lumen formation in tubulogenesis in other systems.
Volume 115(38)
Pages E8873-E8881
Published 2018-9-18
DOI 10.1073/pnas.1808756115
PII 1808756115
PMID 30158171
PMC PMC6156656
MeSH 14-3-3 Proteins / genetics 14-3-3 Proteins / physiology* Animals Benzaldehydes / pharmacology Ciona intestinalis / embryology* Ciona intestinalis / genetics Cytoplasm / metabolism Cytoskeletal Proteins / genetics Cytoskeletal Proteins / metabolism Endothelial Cells / physiology* Larva / growth & development Membrane Proteins / genetics Membrane Proteins / metabolism Microfilament Proteins / genetics Microfilament Proteins / metabolism Morphogenesis / drug effects Morphogenesis / genetics Morphogenesis / physiology* Morpholinos / genetics Myosin Type II / metabolism Notochord / embryology
IF 9.504
Resource
C.intestinalis / (O.japonicus) Wild C. int