Abstract

BackgroundExtracellular cAMP is a key extracellular signaling molecule that regulates aggregation, cell differentiation and morphogenesis during multi-cellular development of the social amoeba Dictyostelium discoideum. This molecule is produced by three different adenylyl cyclases, encoded by the genes acaA, acrA and acgA, expressed at different stages of development and in different structures.Methodology/Principal FindingsThis article describes the characterization of the promoter region of the acaA gene, showing that it is transcribed from three different alternative promoters. The distal promoter, promoter 1, is active during the aggregation process while the more proximal promoters are active in tip-organiser and posterior regions of the structures. A DNA fragment containing the three promoters drove expression to these same regions and similar results were obtained by in situ hybridization. Analyses of mRNA expression by quantitative RT-PCR with specific primers for each of the three transcripts also demonstrated their different temporal patterns of expression.Conclusions/SignificanceThe existence of an aggregation-specific promoter can be associated with the use of cAMP as chemo-attractant molecule, which is specific for some Dictyostelium species. Expression at late developmental stages indicates that adenylyl cyclase A might play a more important role in post-aggregative development than previously considered.

Highlights

  • Intercellular communication plays a central role in multicellular development, coordinating processes such as cell migration, proliferation and differentiation that are the basis for morphogenesis

  • The comparison of the nucleotide sequence of the rapid amplification of the cDNA ends (RACE) products with that of the D. discoideum genome indicated that the acaA gene is transcribed from three different promoter regions distributed along the 4 kb long lsm2-acaA intergenic region

  • The results obtained with the promoter analysis were supported by the in situ hybridization experiments and by the temporal patterns of expression observed for the three acaA transcripts

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Summary

Introduction

Intercellular communication plays a central role in multicellular development, coordinating processes such as cell migration, proliferation and differentiation that are the basis for morphogenesis. One of the simplest organisms where these processes have been studied is the social amoeba Dictyostelium discoideum (for recent reviews, [1,2]). These organisms live as individual amoebe in forest soils. Extracellular cAMP is a key extracellular signaling molecule that regulates aggregation, cell differentiation and morphogenesis during multi-cellular development of the social amoeba Dictyostelium discoideum. This molecule is produced by three different adenylyl cyclases, encoded by the genes acaA, acrA and acgA, expressed at different stages of development and in different structures

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