Co-Immunoprecipitation-Coupled Mass Spectrometry Analysis of Zyxin’s Interactome and Phosphosites in Early Xenopus laevis Development

TL;DR

This study explores how Zyxin protein interacts with other proteins and gets modified during early frog development, revealing its role in controlling cell adhesion and gene activity.

Contribution

The study provides a detailed, stage-specific map of Zyxin’s protein interactions and phosphorylation events during early Xenopus development.

Findings

Zyxin interacts with focal adhesion components, transcriptional regulators, and kinases in a developmentally regulated manner.

Stage-specific phosphorylation of Zyxin isoforms suggests dynamic post-translational control of its functions.

Zyxin acts as a central regulator of cell adhesion, survival, and gene programs during morphogenesis.

Abstract

Protein complexes, assembled by scaffold proteins, act as molecular machines driving development. The mechanosensitive adapter protein Zyxin is a key example, integrating actin cytoskeleton dynamics with gene expression. However, the developmental regulation of its interactions and post-translational modifications remains poorly understood. Here, we characterize the dynamic Zyxin interactome across three early developmental stages of Xenopus laevis (from gastrulation to neurulation) using co-immunoprecipitation coupled with quantitative mass spectrometry (DDA and DIA). We identify stage-specific changes in Zyxin’s association with core focal adhesion components, transcriptional regulators and kinases. Furthermore, we uncover developmentally regulated phosphorylation events on isoforms, suggesting dynamic post-translational control of its interactions. Our work provides a comprehensive resource that positions Zyxin as a central orchestrator of cell adhesion, survival, and gene regulatory programs during morphogenesis. These findings underscore the role of Zyxin as a multifaceted regulatory hub, with important implications for understanding tissue homeostasis and related pathologies.

Figures