Spatially resolved cell polarity proteomics of a human epiblast model
| Autor: | Venkatesha Basrur, Nikola Sekulovski, Chien-Wei Lin, Deborah L. Gumucio, Peng Zou, Linnea E. Taniguchi, Sicong Wang, Alexey I. Nesvizhskii, Craig A. Johnson, Jianping Fu, Kenichiro Taniguchi, Amber E. Carleton, Chari L. Cortez, Ryan F. Townshend, Mara C. Duncan |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0303 health sciences
Multidisciplinary Chemistry Clathrin adaptor complex Morphogenesis SciAdv r-articles Cell Biology Apical membrane Cell biology 03 medical and health sciences 0302 clinical medicine Epiblast Cell polarity embryonic structures Inner cell mass Human embryogenesis 030217 neurology & neurosurgery Research Articles 030304 developmental biology Epithelial polarity Research Article Developmental Biology |
| Zdroj: | Science Advances |
| ISSN: | 2375-2548 |
| Popis: | APEX2-based spatial proteomics identifies cell polarity proteome of hPSC-cysts, a 3D model of human epiblast cavity formation. Critical early steps in human embryonic development include polarization of the inner cell mass, followed by formation of an expanded lumen that will become the epiblast cavity. Recently described three-dimensional (3D) human pluripotent stem cell–derived cyst (hPSC-cyst) structures can replicate these processes. To gain mechanistic insights into the poorly understood machinery involved in epiblast cavity formation, we interrogated the proteomes of apical and basolateral membrane territories in 3D human hPSC-cysts. APEX2-based proximity bioinylation, followed by quantitative mass spectrometry, revealed a variety of proteins without previous annotation to specific membrane subdomains. Functional experiments validated the requirement for several apically enriched proteins in cyst morphogenesis. In particular, we found a key role for the AP-1 clathrin adaptor complex in expanding the apical membrane domains during lumen establishment. These findings highlight the robust power of this proximity labeling approach for discovering novel regulators of epithelial morphogenesis in 3D stem cell–based models. |
| Databáze: | OpenAIRE |
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