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Published December 1, 2014 | Supplemental Material
Journal Article Open

LINKIN, a new transmembrane protein necessary for cell adhesion

Abstract

In epithelial collective migration, leader and follower cells migrate while maintaining cell-cell adhesion and tissue polarity. We have identified a conserved protein and interactors required for maintaining cell adhesion during a simple collective migration in the developing C. elegans male gonad. LINKIN is a previously uncharacterized, transmembrane protein conserved throughout Metazoa. We identified seven atypical FG-GAP domains in the extracellular domain, which potentially folds into a β-propeller structure resembling the α-integrin ligand-binding domain. C. elegans LNKN-1 localizes to the plasma membrane of all gonadal cells, with apical and lateral bias. We identified the LINKIN interactors RUVBL1, RUVBL2, and α-tubulin by using SILAC mass spectrometry on human HEK 293T cells and testing candidates for lnkn-1-like function in C. elegans male gonad. We propose that LINKIN promotes adhesion between neighboring cells through its extracellular domain and regulates microtubule dynamics through RUVBL proteins at its intracellular domain.

Additional Information

© 2014 eLife Sciences Publications. Received: 21 August 2014; Accepted: 28 November 2014; Published: 1 December 2014. We are grateful to Amir Sapir, Hillel Schwartz, and Srimoyee Ghosh for critical reading of the manuscript, and to TFC's postdocs, Xiaoyi Zhaang and Lin Gui, for assistance with IP/Western blot analysis experiments. Caltech protein expression facility expressed and purified LNKN-1 extracellular domain. Mass spectrometry experiments were performed at the Caltech protein exploration laboratory directed by Dr. Sonja Hess and Prof. Ray Deshaies. Wormbase (wormbase.org) was a valuable resource for C. elegans information as was the British Columbia C. elegans Gene Expression Consortium (http://elegans.bcgsc.ca/home/ge_consortium.html) GFP expression pattern database. The monoclonal antibody developed by Frankel, J. and Nelsen, E.M. was obtained from the Developmental Studies Hydridoma Bank, created by the NICHD of the NIH and maintained at the University of Iowa. Nematode strains were provided by the Caenorhabditis Genetics Center, which is funded by the NIH National Center for Research Resources. C.Z.Y. was funded by a CIRM pre-doctoral training grant. P.W.S. in an investigator with the Howard Hughes Medical Institute, which supported this work. Author contributions: MK, T-FC, Conception and design, Acquisition of data, Analysis and interpretation of data, Drafting or revising the article; CZY, JD, Acquisition of data, Analysis and interpretation of data; PWS, Conception and design, Analysis and interpretation of data, Drafting or revising the article.

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