Identification and characterization of Crumbs polarity complex proteins in Caenorhabditis elegans

Victoria G. Castiglioni, João J. Ramalho, Jason R. Kroll, Riccardo Stucchi, Hanna van Beuzekom, Ruben Schmidt, Maarten Altelaar, Mike Boxem*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Crumbs proteins are evolutionarily conserved transmembrane proteins with essential roles in promoting the formation of the apical domain in epithelial cells. The short intracellular tail of Crumbs proteins are known to interact with several proteins, including the scaffolding protein PALS1 (protein associated with LIN7, Stardust in Drosophila). PALS1 in turn binds to a second scaffolding protein PATJ (PALS1-associated tight junction protein) to form the core Crumbs/PALS1/PATJ complex. While essential roles in epithelial organization have been shown for Crumbs proteins in Drosophila and mammalian systems, the three Caenorhabditis elegans crumbs genes are dispensable for epithelial polarization and development. Here, we investigated the presence and function of PALS1 and PATJ orthologs in C. elegans. We identified MAGU-2 as the C. elegans ortholog of PALS1 and show that MAGU-2 interacts with all three Crumbs proteins and localizes to the apical membrane domain of intestinal epithelial cells in a Crumbs-dependent fashion. Similar to crumbs mutants, magu-2 deletion showed no epithelial polarity defects. We also identified MPZ-1 as a candidate ortholog of PATJ based on the physical interaction with MAGU-2 and sequence similarity with PATJ proteins. However, MPZ-1 is not broadly expressed in epithelial tissues and, therefore, not likely a core component of the C. elegans Crumbs complex. Finally, we show overexpression of the Crumbs proteins EAT-20 or CRB-3 can lead to apical membrane expansion in the intestine. Our results shed light on the composition of the C. elegans Crumbs complex and indicate that the role of Crumbs proteins in promoting apical domain formation is conserved.

Original languageEnglish
Article number101786
Pages (from-to)1-16
JournalJournal of Biological Chemistry
Volume298
Issue number4
DOIs
Publication statusPublished - Apr 2022

Keywords

  • C. elegans
  • cell polarity
  • Crb
  • Crumbs
  • epithelial polarity
  • MAGU-2
  • MPZ-1
  • PALS1
  • PATJ
  • Stardust

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