La Trobe

Structural analysis of phosphorylation-associated interactions of human MCC with Scribble PDZ domains

journal contribution
posted on 2025-05-06, 01:55 authored by Sofia Caria, Bryce Stewart, Ruitao Jin, Brian SmithBrian Smith, Patrick HumbertPatrick Humbert, Marc KvansakulMarc Kvansakul
Scribble is a crucial adaptor protein that plays a pivotal role during establishment and control of cell polarity, impacting many physiological processes ranging from cell migration to immunity and organization of tissue architecture. Scribble harbours a leucine-rich repeat domain and four PDZ domains that mediate most of Scribble's interactions with other proteins. It has become increasingly clear that post-translational modifications substantially impact Scribble–ligand interactions, with phosphorylation being a major modulator of binding to Scribble. To better understand how Scribble PDZ domains direct cell polarity signalling and how phosphorylation impacts this process, we investigated human Scribble interactions with MCC (Mutated in Colorectal Cancer). We systematically evaluated the ability of all four individual Scribble PDZ domains to bind the PDZ-binding motif (PBM) of MCC as well as MCC phosphorylated at the −1 Ser position. We show that Scribble PDZ1 and PDZ3 are the major interactors with MCC, and that modifications to Ser at the −1 position in the MCC PBM only has a minor effect on binding to Scribble PDZ domains. We then examined the structural basis for these observations by determining the crystal structures of Scribble PDZ1 domain bound to both the unphosphorylated MCC PBM as well as phosphorylated MCC. Our structures indicated that phospho-Ser at the −1 position in MCC is not involved in major contacts with Scribble PDZ1, and in conjunction with our affinity measurements suggest that the impact of phosphorylation at the −1 position of MCC must extend beyond a simple modulation of the affinity for Scribble PDZ domains.

Funding

This work was supported in whole or part by the National Health and Medical Research Council Australia (Project Grant APP1103871 to MK, POH; Senior Research Fellowship APP1079133 to POH), Australian Research Council (Fellowship FT130101349 to MK) and La Trobe University (Research focus area "Understanding Disease" project grant and Scholarship to BZS). RJ acknowledges receipt of Australian Research Training Scholarships. Part of this work was undertaken using resources from the National Computational Infrastructure, which is supported by the Australian Government and provided through Intersect Australia Ltd, and through the HPC-GPGPU Facility, which was established with the assistance of LIEF Grant LE170100200.

History

Publication Date

2019-12-01

Journal

FEBS Journal

Volume

286

Issue

14

Pagination

16p. (p. 4821-5032)

Publisher

Wiley

ISSN

1742-464X

Rights Statement

© 2019 Federation of European Biochemical Societies

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