@article{ba495b45809f4be0a6db630b14f5d062,
title = "The Wnt/PCP formin Daam1 drives cell-cell adhesion during nephron development",
abstract = "E-cadherin junctions facilitate assembly and disassembly of cell contacts that drive development and homeostasis of epithelial tissues. In this study, using Xenopus embryonic kidney and Madin-Darby canine kidney (MDCK) cells, we investigate the role of the Wnt/planar cell polarity (PCP) formin Daam1 (Dishevelled-associated activator of morphogenesis 1) in regulating E-cadherin-based intercellular adhesion. Using live imaging, we show that Daam1 localizes to newly formed cell contacts in the developing nephron. Furthermore, analyses of junctional filamentous actin (F-actin) upon Daam1 depletion indicate decreased microfilament localization and slowed turnover. We also show that Daam1 is necessary for efficient and timely localization of junctional E-cadherin, mediated by Daam1{\textquoteright}s formin homology domain 2 (FH2). Finally, we establish that Daam1 signaling promotes organized movement of renal cells. This study demonstrates that Daam1 formin junctional activity is critical for epithelial tissue organization.",
keywords = "Daam1, E-cadherin, F-actin, Wnt, Xenopus, adhesion, formin, kidney, nephron, tubulogenesis",
author = "Vanja Krneta-Stankic and Corkins, {Mark E.} and Adriana Paulucci-Holthauzen and Malgorzata Kloc and Gladden, {Andrew B.} and Miller, {Rachel K.}",
note = "Funding Information: We thank the Miller lab, Pierre McCrea, and Jae-II Park lab members for lively discussions and suggestions regarding the manuscript. We also thank Richard Behringer, Yoshihiro Komatsu, Oleh Pochynyuk, and Anna Marie Sokac for help with the project. We thank Kenneth Dunner Jr. at the UT MD Anderson High-Resolution Electron Microscopy Facility and CCSG grant NIH P30CA016672 for supporting the TEM data. We are also thankful to Raymond Habas, Bruce Goode, John Wallingford, Norihiro Sudou, Masanori Taira, Raymond Keller, and William Bement for providing antibodies and constructs. We thank the instructors and teaching assistants of the 2015 National Xenopus Resource Advanced Imaging Course. We are grateful to J.C. Whitney and T.H. Gomez for taking care of the animals. We are grateful to the UTHealth Office of the Executive Vice President and Chief Academic Officer and the Department of Pediatrics Microscopy Core for funding the Zeiss LSM800 confocal microscope. We also thank the BSRB Microscopy Facility in the Department of Genetics, UT-MD Anderson Cancer Center. This work was funded by NIH NIDDK grants (K01DK092320, R03DK118771, and R01DK115655 to R.K.M.); startup funding from the Department of Pediatrics, Pediatric Research Center at the McGovern Medical School (to R.K.M.); The Antje Wuelfrath Gee and Harry Gee, Jr. Family Legacy Scholarship (to V.K.-S.), and The Gigli Family Endowed Scholarships (to V.K.-S.). V.K.-S. conceived the project, performed experiments, analyzed data, and wrote the manuscript. M.E.C. cloned pCS2-mCherry-Daam1 and pCS2-mCherry-Daam1 (Ile698Ala) constructs, performed experiments assessing the effectiveness of Daam1 MO KD at NF stages 39/40, and contributed to data validation. V.K.-S. M.E.C. A.B.G. and R.K.M. performed experiments to generate MDCK shDaam1 cells. V.K.-S. and A.P.-H. imaged the wound healing assays and conducted FRAP experiments. V.K.-S. and M.K. conducted TEM imaging analyses. A.B.G. and R.K.M. supervised the project. All authors were involved in critical evaluation and editing of the manuscript. The authors declare no competing interests. Funding Information: We thank the Miller lab, Pierre McCrea, and Jae-II Park lab members for lively discussions and suggestions regarding the manuscript. We also thank Richard Behringer, Yoshihiro Komatsu, Oleh Pochynyuk, and Anna Marie Sokac for help with the project. We thank Kenneth Dunner Jr. at the UT MD Anderson High-Resolution Electron Microscopy Facility and CCSG grant NIH P30CA016672 for supporting the TEM data. We are also thankful to Raymond Habas, Bruce Goode, John Wallingford, Norihiro Sudou, Masanori Taira, Raymond Keller, and William Bement for providing antibodies and constructs. We thank the instructors and teaching assistants of the 2015 National Xenopus Resource Advanced Imaging Course. We are grateful to J.C. Whitney and T.H. Gomez for taking care of the animals. We are grateful to the UTHealth Office of the Executive Vice President and Chief Academic Officer and the Department of Pediatrics Microscopy Core for funding the Zeiss LSM800 confocal microscope. We also thank the BSRB Microscopy Facility in the Department of Genetics, UT-MD Anderson Cancer Center. This work was funded by NIH NIDDK grants ( K01DK092320 , R03DK118771 , and R01DK115655 to R.K.M.); startup funding from the Department of Pediatrics , Pediatric Research Center at the McGovern Medical School (to R.K.M.); The Antje Wuelfrath Gee and Harry Gee, Jr. Family Legacy Scholarship (to V.K.-S.), and The Gigli Family Endowed Scholarships (to V.K.-S.). Publisher Copyright: {\textcopyright} 2021 The Authors",
year = "2021",
month = jul,
day = "6",
doi = "10.1016/j.celrep.2021.109340",
language = "English (US)",
volume = "36",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "1",
}