Abstract

Cyclic AMP acts as a secondary messenger involving different cellular functions in eukaryotes. Here, proteomic and transcriptomic profiling has been combined to identify novel early developmentally regulated proteins in eukaryote cells. These proteomic and transcriptomic experiments were performed in Dictyostelium discoideum given the unique advantages that this organism offers as a eukaryotic model for cell motility and as a nonmammalian model of human disease. By comparing whole-cell proteome analysis of developed (cAMP-pulsed) wild-type AX2 cells and an independent transcriptomic analysis of developed wild-type AX4 cells, our results show that up to 70% of the identified proteins overlap in the two independent studies. Among them, we have found 26 proteins previously related to cAMP signaling and identified 110 novel proteins involved in calcium signaling, adhesion, actin cytoskeleton, the ubiquitin-proteasome pathway, metabolism, and proteins that previously lacked any annotation. Our study validates previous findings, mostly for the canonical cAMP-pathway, and also generates further insight into the complexity of the transcriptomic changes during early development. This article also compares proteomic data between parental and cells lacking glkA, a GSK-3 kinase implicated in substrate adhesion and chemotaxis in Dictyostelium. This analysis reveals a set of proteins that show differences in expression in the two strains as well as overlapping protein level changes independent of GlkA.

Highlights

  • The soil-dwelling social amoeba Dictyostelium discoideum is an excellent model organism for the study of directed cell migration since Dictyostelium cells show robust chemotactic responses to the chemoattractant cAMP [1,2,3,4]

  • We have identified a proteomic and transcriptomic signature of early developmentally regulated proteins, including 26 proteins previously related to cAMP signaling and 110 novel proteins involved in calcium signaling, adhesion, actin cytoskeleton, the ubiquitin-proteasome pathway, metabolism, and proteins that previously lacked any annotation

  • AX4 strains have some genomic and behavioural differences and there are likely to be strain differences under the experimental conditions analyzed in this study, we argue that any changes shared between the two strains provide with robust data

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Summary

Introduction

The soil-dwelling social amoeba Dictyostelium discoideum is an excellent model organism for the study of directed cell migration since Dictyostelium cells show robust chemotactic responses to the chemoattractant cAMP [1,2,3,4]. Cell migration is fundamental to establishing and maintaining the proper organization of multicellular organisms, from large-scale migrations of epithelial sheets during gastrulation, to the movement of individual cells during development of the nervous system [5,6,7]. Cell migration is essential for proper immune response [8], wound repair [9], and tissue homeostasis [10], while aberrant cell migration is found in various pathologies [7]. A directional sensing system biases pseudopodia formation towards the source of the chemoattractant, and orients

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