Mef2A, a homologue of animal Mef2 transcription factors, regulates cell differentiation in Dictyostelium discoideum

BMC Developmental Biology

Leandro Sastre¹  Teresa Suarez²  Irene Fernandez-Aguado¹  María Galardi-Castilla¹ 

[1] Instituto de Investigaciones Biomédicas de Madrid (Biomedical Research Institute of Madrid), CSIC/UAM, C/Arturo Duperier 4, 28029 Madrid, Spain;Centro de Investigaciones Biologicas (Biological Research Center), CSIC, C/Ramiro de Maeztu, 9, 28040, Madrid, Spain

关键词: Transcription factor;    Cell-type patterning;    Development;    Cell differentiation;    Dictyostelium;    Myocyte enhancer factor 2;   

Others  :  1085945
DOI  :  10.1186/1471-213X-13-12

 received in 2012-11-14, accepted in 2013-04-05,  发布年份 2013

【 摘 要 】

Background

Transcription factors from the MADS-box family play a relevant role in cell differentiation and development and include the animal SRF (serum response factor) and MEF2 (myocyte enhancer factor 2) proteins. The social amoeba Dictyostelium discoideum contains four genes coding for MADS-box transcription factors, two of these genes code for proteins that are more similar to SRF, and the other two code for proteins that are more similar to MEF2 animal factors.

Results

The biological function of one of the two genes that codes for MEF2-related proteins, a gene known as mef2A, is described in this article. This gene is expressed under the transcriptional control of two alternative promoters in growing cells, and its expression is induced during development in prespore cells. Mutant strains where the mef2A gene has been partially deleted were generated to study its biological function. The mutant strains showed reduced growth when feeding on bacteria and were able to develop and form fruiting bodies, but spore production was significantly reduced. A study of developmental markers showed that prespore cells differentiation was impaired in the mutant strains. When mutant and wild-type cells were set to develop in chimeras, mutant spores were underrepresented in the fruiting bodies. The mutant cells were also unable to form spores in vitro. In addition, mutant cells also showed a poor contribution to the formation of the tip-organizer and the upper region of slugs and culminant structures. In agreement with these observations, a comparison of the genes transcribed by mutant and wild-type strains during development indicated that prestalk gene expression was enhanced, while prespore gene expression decreased in the mef2A^- strain.

Conclusions

Our data shows that mef2A plays a role in cell differentiation in D. discoideum and modulates the expression of prespore and prestalk genes.

【 授权许可】

   
2013 Galardi-Castilla et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

Figure 6.

Figure 7.

Figure 1.

【 参考文献 】

• [1]Theiben G, Kim JT, Saedler H: Classification and Phylogeny of the MADS-box Multigene Family Suggest Defined Roles of MADS-box Gene Subfamilies in the Morphological Evolution of Eukaryotes. J mol evol 1996, 43:484-516.
• [2]Gramzow L, Ritz MS, Theissen G: On the origin of MADS-domain transcription factors. Trends Genet 2010, 26(4):149-153.
• [3]Shore P, Sharrocks AD: The MADS-box family of transcription factors. Eur J Biochem 1995, 229(1):1-13.
• [4]Treisman R: DNA-binding proteins. Inside the MADS box. Nature 1995, 376(6540):468-469.
• [5]Becker A, Theissen G: The major clades of MADS-box genes and their role in the development and evolution of flowering plants. Mol Phylogenet Evol 2003, 29(3):464-489.
• [6]Messenguy F, Dubois E: Role of MADS box proteins and their cofactors in combinatorial control of gene expression and cell development. Gene 2003, 316:1-21.
• [7]Pollock R, Treisman R: A sensitive method for the determination of protein-DNA binding specificities. Nucleic Acids Res 1990, 18(21):6197-6204.
• [8]Sun Q, Chen G, Streb JW, Long X, Yang Y, Stoeckert CJ Jr, Miano JM: Defining the mammalian CArGome. Genome Res 2007, 16:197-207.
• [9]Black BL, Olson EN: Transcriptional control of muscle development by myocyte enhancer factor-2 (MEF2) proteins. Annu Rev Cell Dev Biol 1998, 14:167-196.
• [10]Treisman R, Ammerer G: The SRF and MCM1 transcription factors. Curr Opin Genet Dev 1992, 2(2):221-226.
• [11]Chang VK, Donato JJ, Chan CS, Tye BK: Mcm1 promotes replication initiation by binding specific elements at replication origins. Mol Cell Biol 2004, 24:6514-6524.
• [12]Schratt G, Philippar U, Berger J, Schwarz H, Heidenreich O, Nordheim A: Serum response factor is crucial for actin cytoskeletal organization and focal adhesion assembly in embryonic stem cells. J Cell Biol 2002, 156:737-750.
• [13]Arsenian S, Weinhold B, Oelgeschlager M, Ruther U, Nordheim A: Serum response factor is essential for mesoderm formation during mouse embryogenesis. Embo J 1998, 17(21):6289-6299.
• [14]Miano JM: Role of serum response factor in the pathogenesis of disease. Lab Invest 2010, 90(9):1274-1284.
• [15]Miano JM, Long X, Fujiwara K: Serum response factor: master regulator of the actin cytoskeleton and contractil apparatus. Am J Physiol Cell Physiol 2007, 292:C70-C81.
• [16]Olson EN, Nordheim A: Linking actin dynamics and gene transcription to drive cellular motile functions. Nat Rev Mol Cell Biol 2010, 11(5):353-365.
• [17]Sandmann T, Jensen LJ, Jakobsen JS, Karzynski MM, Eichenlaub MP, Bork P, Furlong EE: A temporal map of transcription factor activity: mef2 directly regulates target genes at all stages of muscle development. Dev Cell 2006, 10(6):797-807.
• [18]Naya FJ, Olson E: MEF2: a transcriptional target for signaling pathways controlling skeletal muscle growth and differentiation. Curr Opin Cell Biol 1999, 11(6):683-688.
• [19]Molkentin JD, Olson EN: Combinatorial control of muscle development by basic helix-loop-helix and MADS-box transcription factors. Proc Natl Acad Sci U S A 1996, 93(18):9366-9373.
• [20]McKinsey TA, Zhang CL, Olson EN: MEF2: a calcium-dependent regulator of cell division, differentiation and death. Trends Biochem Sci 2002, 27(1):40-47.
• [21]Lin Q, Schwarz J, Bucana C, Olson EN: Control of mouse cardiac morphogenesis and myogenesis by transcription factor MEF2C. Science 1997, 276(5317):1404-1407.
• [22]Naya FJ, Black BL, Wu H, Bassel-Duby R, Richardson JA, Hill JA, Olson EN: Mitochondrial deficiency and cardiac sudden death in mice lacking the MEF2A transcription factor. Nat Med 2002, 8(11):1303-1309.
• [23]Potthoff M, Olson EN: MEF2: a central regulator of diverse developmental programs. Development 2007, 134:4131-4140.
• [24]Arnold MA, Kim Y, Czubryt MP, Phan D, McAnally J, Qi X, Shelton JM, Richardson JA, Bassel-Duby R, Olson EN: MEF2C transcription factor controls chondrocyte hypertrophy and bone development. Dev Cell 2007, 12(3):377-389.
• [25]Williams JG: Dictyostelium finds new roles to model. Genetics 2010, 185(3):717-726.
• [26]Schaap P: Evolutionary crossroads in developmental biology: Dictyostelium discoideum. Development 2011, 138(3):387-396.
• [27]Urushihara H: Social amoeba and the origin of multicellularity. Dev Growth Differ 2011, 53(4):451.
• [28]Escalante R, Sastre L: A serum response factor homolog is required for spore differentiation in Dictyostelium. Development 1998, 125:3801-3808.
• [29]Escalante R, Yamada Y, Cotter D, Sastre L, Sameshima M: The MADS-box transcription factor SrfA is required for actin cytoskeleton organization and spore coat stability during Dictyostelium sporulation. Mechanisms of Development 2004, 121(1):51-56.
• [30]Galardi-Castilla M, Pergolizzi B, Bloomfield G, Skelton J, Ivens A, Kay RR, Bozzaro S, Sastre L: SrfB, a member of the Serum Response Factor family of transcription factors, regulates starvation response and early development in Dictyostelium. Dev Biol 2008, 316(2):260-274.
• [31]Williams JG: Transcriptional regulation of Dictyostelium pattern formation. EMBO Rep 2006, 7(7):694-698.
• [32]Kay RR: Evidence that elevated intracellular cyclic AMP triggers spore maturation in Dictyostelium. Development 1989, 105:753-759.
• [33]Shimada N, Kanno-Tanabe N, Minemura K, Kawata T: GBF-dependent family genes morphologically suppress the partially active Dictyostelium STATa strain. Dev Genes Evol 2008, 218(2):55-68.
• [34]Guo K, Anjard C, Harwood A, Kim HJ, Newell PC, Gross JD: A myb-related protein required for culmination in Dictyostelium. Development 1999, 126:2813-2822.
• [35]Sillo A, Bloomfield G, Balest A, Balbo A, Pergolizzi B, Peracino B, Skelton J, Ivens A, Bozzaro S: Genome-wide transcriptional changes induced by phagocytosis or growth on bacteria in Dictyostelium. BMC Genomics 2008, 9:291. BioMed Central Full Text
• [36]Jermyn KA, Berks M, Kay RR, Williams JG: Two distinct classes of prestalk-enriched mRNA sequences in Dictyostelium discoideum. Development 1987, 100:745-755.
• [37]Benabentos R, Hirose S, Sucgang R, Curk T, Katoh M, Ostrowski EA, Strassmann JE, Queller DC, Zupan B, Shaulsky G: Polymorphic members of the lag gene family mediate kin discrimination in Dictyostelium. Curr Biol 2009, 19(7):567-572.
• [38]Parkinson K, Bolourani P, Traynor D, Aldren NL, Kay RR, Weeks G, Thompson CR: Regulation of Rap1 activity is required for differential adhesion, cell-type patterning and morphogenesis in Dictyostelium. J Cell Sci 2009, 122(Pt 3):335-344.
• [39]Sawarkar R, Visweswariah SS, Nellen W, Nanjundiah V: Histone deacetylases regulate multicellular development in the social amoeba Dictyostelium discoideum. J Mol Biol 2009, 391(5):833-848.
• [40]Dubin MJ, Kasten S, Nellen W: Characterization of the Dictyostelium homolog of chromatin binding protein DET1 suggests a conserved pathway regulating cell type specification and developmental plasticity. Eukaryot Cell 2011, 10(3):352-362.
• [41]MacWilliams H, Doquang K, Pedrola R, Dollman G, Grassi D, Peis T, Tsang A, Ceccarelli A: A retinoblastoma ortholog controls stalk/spore preference in Dictyostelium. Development 2006, 133(7):1287-1297.
• [42]Miska EA, Karlsson C, Langley E, Nielsen SJ, Pines J, Kouzarides T: HDAC4 deacetylase associates with and represses the MEF2 transcription factor. EMBO J 1999, 18(18):5099-5107.
• [43]Han J, Jiang Y, Li Z, Kravchenko VV, Ulevitch RJ: Activation of the transcription factor MEF2C by the MAP kinase p38 in inflammation. Nature 1997, 386(6622):296-299.
• [44]Nguyen HN, Hadwiger JA: The Galpha4 G protein subunit interacts with the MAP kinase ERK2 using a D-motif that regulates developmental morphogenesis in Dictyostelium. Dev Biol 2009, 335(2):385-395.
• [45]Nguyen HN, Raisley B, Hadwiger JA: MAP kinases have different functions in Dictyostelium G protein-mediated signaling. Cell Signal 2010, 22(5):836-847.
• [46]Mann SKO, Richardson DL, Lee S, Kimmel AR, Firtel RA: Expression of cAMP-dependent protein kinase in prespore cells is sufficient to induce spore cell differentiation in Dictyostelium. Proc Natl Acad Sci USA 1994, 91:10561-10565.
• [47]Loomis WF, Shaulsky G: Developmental changes in transcriptional profiles. Dev Growth Differ 53(4):567-575.
• [48]Maruo T, Sakamoto H, Iranfar N, Fuller D, Morio T, Urushihara H, Tanaka Y, Maeda M, Loomis WF: Control of cell type proportioning in Dictyostelium discoideum by differentiation-inducing factor as determined by in sity hybridization. Euk Cell 2004, 3(5):1241-1248.
• [49]Ho Sui SJ, Mortimer JR, Arenillas DJ, Brumm J, Walsh CJ, Kennedy BP, Wasserman WW: oPOSSUM: identification of over-represented transcription factor binding sites in co-expressed genes. Nucleic Acids Res 2005, 33(10):3154-3164.
• [50]Chen G, Hata N, Zhang MQ: Transcription factor binding element detection using functional clustering of mutant expression data. Nucleic Acids Res 2004, 32(8):2362-2371.
• [51]Escalante R, Vicente JJ, Moreno N, Sastre L: The MADS-box gene srfA is expressed in a complex pattern under the control of alternative promoters and is essential for different aspects of Dictyostelium development. Dev Biol 2001, 235(2):314-329.
• [52]Faure M, Franke J, Hall AL, Podgorski GJ, Kessin RH: The cyclic nucleotide phosphodiesterase gene of Dictyostelium discoideum contains 3 promoters specific for growth, aggregation, and late development. Mol Cell Biol 1990, 10:1921-1930.
• [53]Louis JM, III Saxe CL, Kimmel AR: Two transmembrane signaling mechanisms control expression of the cAMP receptor gene CAR1 during Dictyostelium development. Proc Natl Acad Sci USA 1993, 90:5969-5973.
• [54]Galardi-Castilla M, Garciandia A, Suarez T, Sastre L: The Dictyostelium discoideum acaA gene is transcribed from alternative promoters during aggregation and multicellular development. PLoS One 2010, 5(10):e13286.
• [55]Pang KM, Lynes MA, Knecht DA: Variables controlling the expression level of exogenous genes in Dictyostelium. Plasmid 1999, 41:187-197.
• [56]Adachi H, Hasebe T, Yoshinaga K, Ohta T, Sutoh K: Isolation of Dictyostelium discoideum cytokinesis mutants by restriction enzyme-mediated integration of the blasticidin S resistance marker. Biochem Biophys Res Commun 1994, 205:1808-1814.
• [57]Shaulsky G, Loomis WF: Cell type regulation in response to expression of ricin-A in Dictyostelium. Dev Biol 1993, 160:85-98.
• [58]Thompson J, Gibson T, Plewniak F, Jeanmougins F, Higgins D: The ClustalX windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucl Acids Res 1997, 25:4876-4882.
• [59]Saitou N, Nei M: The Neighbor-Joining method for reconstructing phylogenetic trees. Mol Biol Evol 1987, 4:406-425.
• [60]Faix J, Kreppel L, Shaulsky G, Schleicher M, Kimmel AR: A rapid and efficient method to generate multiple gene disruptions in Dictyostelium discoideum using a single selectable marker and the Cre-loxP system. Nucleic Acids Res 2004, 32(19):e143.
• [61]Detterbeck S, Morandini P, Wetterauer B, Bachmair A, Fischer K, MacWilliams HK: The ‘prespore-like cells’ of Dictyostelium have ceased to express a prespore gene: Analysis using short-lived beta-galactosidases as reporters. Development 1994, 120:2847-2855.
• [62]Jermyn KA, Williams JG: An analysis of culmination in Dictyostelium using prestalk and stalk-specific cell autonomous markers. Development 1991, 111:779-787.
• [63]Meima ME, Weening KE, Schaap P: Vectors for expression of proteins with single or combinatorial fluorescent protein and tandem affinity purification tags in Dictyostelium. Protein Expr Purif 2007, 53(2):283-288.
• [64]Escalante R, Sastre L: Investigating gene expression: In situ hybridization and reporter genes. In Dictyostelium discoideum protocols. vol. 346. Edited by Eichinger L, Rivero F. Totowa, NJ: Humana Press; 2006:230-247.
• [65]Serafimidis I, Kay RR: New prestalk and prespore inducing signals in Dictyostelium. Dev Biol 2005, 282(2):432-441.
• [66]Trapnell C, Pachter L, Salzberg SL: TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 2009, 25(9):1105-1111.
• [67]Anders S, Huber W: Differential expression analysis for sequence count data. Genome Biol 2010, 11(10):106. BioMed Central Full Text