The prokaryote messenger c-di-GMP triggers stalk cell differentiation in Dictyostelium


Cyclic di-(3′:5′)-guanosine monophosphate (c-di-GMP) is a major prokaryote signalling intermediate that is synthesized by diguanylate cyclases and triggers sessility and biofilm formation1,2. We detected the first eukaryote diguanylate cyclases in all major groups of Dictyostelia. On food depletion, Dictyostelium discoideum amoebas collect into aggregates, which first transform into migrating slugs and then into sessile fruiting structures3. These structures consist of a spherical spore mass that is supported by a column of stalk cells and a basal disk. A polyketide, DIF-1, which induces stalk-like cells in vitro, was isolated earlier4. However, its role in vivo proved recently to be restricted to basal disk formation5. Here we show that the Dictyostelium diguanylate cyclase, DgcA, produces c-di-GMP as the morphogen responsible for stalk cell differentiation. Dictyostelium discoideum DgcA synthesized c-di-GMP in a GTP-dependent manner and was expressed at the slug tip, which is the site of stalk cell differentiation. Disruption of the DgcA gene blocked the transition from slug migration to fructification and the expression of stalk genes. Fructification and stalk formation were restored by exposing DgcA-null slugs to wild-type secretion products or to c-di-GMP. Moreover, c-di-GMP, but not cyclic di-(3′:5′)-adenosine monophosphate, induced stalk gene expression in dilute cell monolayers. Apart from identifying the long-elusive stalk-inducing morphogen, our work also identifies a role for c-di-GMP in eukaryotes.

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Figure 1: Identification and disruption of DGCs.
Figure 2: DgcA expression pattern.
Figure 3: Biological role of c-di-GMP.
Figure 4: Bioassay of DGC activity.

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Primary accessions


Data deposits

DNAsequences for D. lacteumandA. subglobosumDgcAhave been submitted to Genbank under accession numbers JQ676836 and JQ676837, respectively.


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We thank R. R. Kay for alerting us to the presence of a putative diguanylate cyclase in the D. discoideum genome. U. Jenal is gratefully acknowledged for advice and an EAL-PDE construct in an early phase of the project. We are grateful to the late H. MacWilliams for plasmids PspA-ile-gal, EcmA-ile-gal and PstO-ile-gal, and to C.Thompson for dmta cells. We thank W. Chen and D. Lamont for mass spectrometry and C. Sugden for guidance with qRT–PCR. We are grateful to H. Urushihara and the A. subglobosum genome project ( = as) for the A. subglobosum DgcA sequence. This research was supported by Wellcome Trust Project grant 090276 and BBSRC grant BB/G020426.

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Z.C. and P.S. designed the experiments and wrote the manuscript. Z.C. performed the experiments.

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Correspondence to Pauline Schaap.

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The authors declare no competing financial interests.

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This file contains Supplementary Figures 1-7, Supplementary Table 1 and additional references. (PDF 4943 kb)

This movie file shows the developmental phenotype of the diguanylate cyclase null mtutant. (MOV 11651 kb)

Supplementary Movie 1

This movie file shows the developmental phenotype of the diguanylate cyclase null mtutant. (MOV 11651 kb)

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Chen, Z., Schaap, P. The prokaryote messenger c-di-GMP triggers stalk cell differentiation in Dictyostelium. Nature 488, 680–683 (2012).

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