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

Abstract

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.

Accession codes

Primary accessions

GenBank/EMBL/DDBJ

Data deposits

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

References

  1. 1

    Jenal, U. &. Malone, J. Mechanisms of cyclic-di-GMP signaling in bacteria. Annu. Rev. Genet. 40, 385–407 (2006)

    CAS  Article  Google Scholar 

  2. 2

    Hengge, R. Principles of c-di-GMP signalling in bacteria. Nature Rev. Microbiol. 7, 263–273 (2009)

    CAS  Article  Google Scholar 

  3. 3

    Schaap, P. The evolution of size and pattern in the social amoebas. Bioessays 29, 635–644 (2007)

    CAS  Article  Google Scholar 

  4. 4

    Morris, H. R., Taylor, G. W., Masento, M. S., Jermyn, K. A. & Kay, R. R. Chemical structure of the morphogen differentiation inducing factor from Dictyostelium discoideum. Nature 328, 811–814 (1987)

    ADS  CAS  Article  Google Scholar 

  5. 5

    Saito, T., Kato, A. & Kay, R. R. DIF-1 induces the basal disc of the Dictyostelium fruiting body. Dev. Biol. 317, 444–453 (2008)

    CAS  Article  Google Scholar 

  6. 6

    Eichinger, L. et al. The genome of the social amoeba Dictyostelium discoideum. Nature 435, 43–57 (2005)

    ADS  CAS  Article  Google Scholar 

  7. 7

    Schaap, P. et al. Molecular phylogeny and evolution of morphology in the social amoebas. Science 314, 661–663 (2006)

    ADS  CAS  Article  Google Scholar 

  8. 8

    Chan, C. et al. Structural basis of activity and allosteric control of diguanylate cyclase. Proc. Natl Acad. Sci. USA 101, 17084–17089 (2004)

    ADS  CAS  Article  Google Scholar 

  9. 9

    Raper, K. B. & Fennell, D. I. Stalk formation in Dictyostelium. Bull. Torrey Botan. Club 79, 25–51 (1952)

    Article  Google Scholar 

  10. 10

    Ross, P. et al. Regulation of cellulose synthesis in Acetobacter xylinum by cyclic diguanylic acid. Nature 325, 279–281 (1987)

    ADS  CAS  Article  Google Scholar 

  11. 11

    Blanton, R. L., Fuller, D., Iranfar, N., Grimson, M. J. & Loomis, W. F. The cellulose synthase gene of Dictyostelium. Proc. Natl Acad. Sci. USA 97, 2391–2396 (2000)

    ADS  CAS  Article  Google Scholar 

  12. 12

    Early, A. E. et al. Structural characterization of Dictyostelium discoideum prespore- specific gene D19 and of its product, cell surface glycoprotein PsA. Mol. Cell. Biol. 8, 3458–3466 (1988)

    CAS  Article  Google Scholar 

  13. 13

    Williams, J. G. Transcriptional regulation of Dictyostelium pattern formation. EMBO Rep. 7, 694–698 (2006)

    CAS  Article  Google Scholar 

  14. 14

    Richardson, D. L. & Loomis, W. F. Disruption of the sporulation-specific gene spiA in Dictyostelium discoideum leads to spore instability. Genes Dev. 6, 1058–1070 (1992)

    CAS  Article  Google Scholar 

  15. 15

    Ceccarelli, A., Zhukovskaya, N., Kawata, T., Bozzaro, S. & Williams, J. Characterisation of a DNA sequence element that directs Dictyostelium stalk cell-specific gene expression. Differentiation 66, 189–196 (2000)

    CAS  Article  Google Scholar 

  16. 16

    Ceccarelli, A., Mahbubani, H. & Williams, J. G. Positively and negatively acting signals regulating stalk cell and anterior-like cell differentiation in Dictyostelium. Cell 65, 983–989 (1991)

    CAS  Article  Google Scholar 

  17. 17

    Thompson, C. R. & Kay, R. R. The role of DIF-1 signaling in Dictyostelium development. Mol. Cell 6, 1509–1514 (2000)

    CAS  Article  Google Scholar 

  18. 18

    Brookman, J. J., Town, C. D., Jermyn, K. A. & Kay, R. R. Developmental regulation of stalk cell differentiation-inducing factor in Dictyostelium discoideum. Dev. Biol. 91, 191–196 (1982)

    CAS  Article  Google Scholar 

  19. 19

    Wang, M., Van Driel, R. & Schaap, P. Cyclic AMP-phosphodiesterase induces dedifferentiation of prespore cells in Dictyostelium discoideum slugs: evidence that cyclic AMP is the morphogenetic signal for prespore differentiation. Development 103, 611–618 (1988)

    CAS  Google Scholar 

  20. 20

    Shadwick, L. L., Spiegel, F. W., Shadwick, J. D., Brown, M. W. & Silberman, J. D. Eumycetozoa = Amoebozoa?: SSUrDNA phylogeny of protosteloid slime molds and its significance for the amoebozoan supergroup. PLoS ONE 4, e6754 (2009)

    ADS  Article  Google Scholar 

  21. 21

    Meima, M. E., Biondi, R. M. & Schaap, P. Identification of a novel type of cGMP phosphodiesterase that is defective in the chemotactic stmF mutants. Mol. Biol. Cell 13, 3870–3877 (2002)

    CAS  Article  Google Scholar 

  22. 22

    Harwood, A. J. & Drury, L. New vectors for expression of the E. coli lacZ gene in Dictyostelium.. Nucleic Acids Res. 18, 4292 (1990)

    CAS  Article  Google Scholar 

  23. 23

    Dingermann, T. et al. Optimization and in situ detection of Escherichia coli beta-galactosidase gene expression in Dictyostelium discoideum. Gene 85, 353–362 (1989)

    CAS  Article  Google Scholar 

  24. 24

    Williams, J. G. et al. Direct induction of Dictyostelium prestalk gene expression by DIF provides evidence that DIF is a morphogen. Cell 49, 185–192 (1987)

    CAS  Article  Google Scholar 

  25. 25

    Schaap, P. et al. Cell-permeable non-hydrolyzable cAMP derivatives as tools for analysis of signaling pathways controlling gene regulation in Dictyostelium. J. Biol. Chem. 268, 6323–6331 (1993)

    CAS  PubMed  Google Scholar 

  26. 26

    Paul, R. et al. Cell cycle-dependent dynamic localization of a bacterial response regulator with a novel di-guanylate cyclase output domain. Genes Dev. 18, 715–727 (2004)

    CAS  Article  Google Scholar 

  27. 27

    Heidel, A. et al. Phylogeny-wide analysis of social amoeba genomes highlights ancient origins for complex intercellular communication. Genome Res. 21, 1882–1891 (2011)

    CAS  Article  Google Scholar 

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Acknowledgements

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 (http://acytodb.biol.tsukuba.ac.jp/cgi-bin/index.cgi?org = 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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Supplementary Information

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

41586_2012_BFnature11313_MOESM27_ESM.mov

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). https://doi.org/10.1038/nature11313

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