Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published November 13, 2018 | Supplemental Material + Published
Journal Article Open

Discs large 1 controls daughter-cell polarity after cytokinesis in vertebrate morphogenesis

Abstract

Vertebrate embryogenesis and organogenesis are driven by cell biological processes, ranging from mitosis and migration to changes in cell size and polarity, but their control and causal relationships are not fully defined. Here, we use the developing limb skeleton to better define the relationships between mitosis and cell polarity. We combine protein-tagging and -perturbation reagents with advanced in vivo imaging to assess the role of Discs large 1 (Dlg1), a membrane-associated scaffolding protein, in mediating the spatiotemporal relationship between cytokinesis and cell polarity. Our results reveal that Dlg1 is enriched at the midbody during cytokinesis and that its multimerization is essential for the normal polarity of daughter cells. Defects in this process alter tissue dimensions without impacting other cellular processes. Our results extend the conventional view that division orientation is established at metaphase and anaphase and suggest that multiple mechanisms act at distinct phases of the cell cycle to transmit cell polarity. The approach employed can be used in other systems, as it offers a robust means to follow and to eliminate protein function and extends the Phasor approach for studying in vivo protein interactions by frequency-domain fluorescence lifetime imaging microscopy of Förster resonance energy transfer (FLIM-FRET) to organotypic explant culture.

Additional Information

© 2018 the Author(s). Published under the PNAS license. Edited by Richard M. Harland, University of California, Berkeley, CA, and approved September 26, 2018 (received for review August 31, 2017). We thank Prof. Francesca Mariani for her critical reading of the manuscript, Prof. Cheng-Ming Chuong for sharing equipment, Ang Li for statistical analyses and sharing reagents, Francesco Cutrale for helpful discussions and editing, Prof. Rich Roberts for the mRNA display without which FingRs would not exist, and Annie Moradian and Michael Sweredoski of the Proteome Exploration Laboratory at the California Institute of Technology for services, troubleshooting, and experimental advice regarding mass spectroscopy. This study was supported by the University of Southern California Translational Imaging Center. Y.L. and J.A.J. contributed equally to this work. Author contributions: Y.L., J.A.J., and S.E.F. designed research; Y.L., J.A.J., and C.A. performed research; J.H.M. provided all mouse tissues; Y.L., J.A.J., C.A., G.G.G., J.H.M., R.M., R.W.R., D.B.A., and S.E.F. contributed new reagents/analytic tools; Y.L., J.A.J., C.A., and S.E.F. analyzed data; and Y.L., J.A.J., C.A., D.B.A., and S.E.F. wrote the paper. The authors declare no conflict of interest. This article is a PNAS Direct Submission. This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10.1073/pnas.1713959115/-/DCSupplemental.

Attached Files

Published - E10859.full.pdf

Supplemental Material - pnas.1713959115.sapp.pdf

Supplemental Material - pnas.1713959115.sd01.xlsx

Supplemental Material - pnas.1713959115.sm01.avi

Supplemental Material - pnas.1713959115.sm02.avi

Supplemental Material - pnas.1713959115.sm03.avi

Supplemental Material - pnas.1713959115.sm04.avi

Supplemental Material - pnas.1713959115.sm05.avi

Supplemental Material - pnas.1713959115.sm06.avi

Files

pnas.1713959115.sapp.pdf
Files (86.4 MB)
Name Size Download all
md5:00c5b8b13561cb2852cc181eaab67b9b
82.2 MB Preview Download
md5:61dd27700238e4e7ab9d283ed238b6a6
216.3 kB Download
md5:e9e390951d971da8533cc33976b246c5
138.4 kB Download
md5:c54fbec505d2682f0149ce38bff4c8b0
135.5 kB Download
md5:a55c696e9baf90eb5eae8c6811ae7b94
163.9 kB Download
md5:f78c0555429a89e6e0aeaed2134bd51a
306.9 kB Download
md5:b92c015a82a9690752c5c61e381ca814
128.1 kB Download
md5:ca81c3e942715b5df24f3dc15ae9bb0a
3.0 MB Preview Download
md5:80f7e8ad2a63154825628a01b41f2bdb
139.3 kB Download

Additional details

Created:
August 22, 2023
Modified:
October 18, 2023