Structure of Crumbs tail in complex with the PALS1 PDZ-SH3-GK tandem reveals a highly specific assembly mechanism for the apical Crumbs complex

Youjun Li, Zhiyi Wei, Yan Yan, Qingwen Wan, Quansheng Du, Mingjie Zhang

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

The Crumbs (Crb) complex, formed by Crb, PALS1, and PATJ, is evolutionarily conserved in metazoans and acts as a master cellgrowth and -polarity regulator at the apical membranes in polarized epithelia. Crb intracellular functions, including its direct binding to PALS1, are mediated by Crb's highly conserved 37-residue cytoplasmic tail. However, themechanistic basis governing the highly specific Crb-PALS1 complex formation is unclear, as reported interaction between the Crb tail (Crb-CT) and PALS1 PSD-95/DLG/ZO-1 (PDZ) domain is weak and promiscuous. Here we have discovered that the PDZ-Src homolgy 3 (SH3)-Guanylate kinase (GK) tandem of PALS1 binds to Crb-CT with a dissociation constant of 70 nM, which is ∼100-fold stronger than the PALS1 PDZ-Crb-CT interaction. The crystal structure of the PALS1 PDZ-SH3-GK-Crb-CT complex reveals that PDZ-SH3-GK forms a structural supramodulewith all three domains contributing to the tight binding to Crb. Mutations disrupting the tertiary interactions of the PDZ-SH3-GK supramodule weaken the PALS1-Crb interaction and compromise PALS1-mediated polarity establishment in Madin- Darby canine kidney (MDCK) cysts. We further show that specific target binding of other members of membrane-associated guanylate kinases (MAGUKs) (e.g., CASK binding to neurexin) also requires the presence of their PDZ-SH3-GK tandems.

Original languageEnglish (US)
Pages (from-to)17444-17449
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume111
Issue number49
DOIs
StatePublished - Dec 9 2014

Fingerprint

Guanylate Kinases
Canidae
Cysts
Epithelium
Kidney
Mutation
Membranes

Keywords

  • Cell polarity
  • MAGUK
  • PBM
  • Stardust
  • Supramodule

ASJC Scopus subject areas

  • General

Cite this

Structure of Crumbs tail in complex with the PALS1 PDZ-SH3-GK tandem reveals a highly specific assembly mechanism for the apical Crumbs complex. / Li, Youjun; Wei, Zhiyi; Yan, Yan; Wan, Qingwen; Du, Quansheng; Zhang, Mingjie.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 111, No. 49, 09.12.2014, p. 17444-17449.

Research output: Contribution to journalArticle

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