Primary structure and regulation of vegetative specific genes of<i>Dictyostelium discoideum</i>

Singleton, Charles K.; Manning, Suzanne; Ken, Ruey

doi:10.1093/nar/17.23.9679

Cited by 47 publications

(26 citation statements)

References 28 publications

(27 reference statements)

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“…Sequence comparison of EDF-1 by the BLASTX program revealed 66% identity at the amino acid level to MBF1 (multiprotein-bridging factor 1) of the silkworm Bombyx mori, a transcriptional coactivator that may play a role in differentiation (17), and 46% identity to H7, a developmentally regulated gene of Dictyostelium discoideum (18,19). In Fig.…”

Section: Resultsmentioning

confidence: 99%

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EDF-1, a Novel Gene Product Down-regulated in Human Endothelial Cell Differentiation

Dragoni¹,

Mariotti²,

Consalez³

et al. 1998

Journal of Biological Chemistry

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Endothelial cell differentiation is a crucial step in angiogenesis. Here we report the identification of EDF-1, a novel gene product that is down-regulated when endothelial cells are induced to differentiate in vitro. The cDNA encoding EDF-1 was isolated by RNA fingerprinting from human endothelial cells exposed to human immunodeficiency virus type 1 Tat, a viral protein known to be angiogenic. The deduced amino acid sequence of EDF-1 encodes a basic intracellular protein of 148 amino acids that is homologous to MBF1 (multiprotein-bridging factor 1) of the silkworm Bombyx mori and to H7, which is implicated in the early developmental events of Dictyostelium discoideum. Interestingly, human immunodeficiency virus type 1 Tat, which affects endothelial functions, and the phorbol ester 12-Otetradecanoylphorbol-13-acetate and culture on fibrin gels, which promote endothelial differentiation in vitro, all down-regulate EDF-1 expression both at the RNA and protein levels. In addition, the inhibition of EDF-1 translation by an antisense anti-EDF-1 construct results in the inhibition of endothelial cell growth and in the transition from a nonpolar cobblestone phenotype to a polar fibroblast-like phenotype. These data suggest that EDF-1 may play a role in the regulation of human endothelial cell differentiation.

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Section: Resultsmentioning

confidence: 99%

“…Cells-The possibility that EDF-1 is modulated in endothelial differentiation is suggested by the similarity between EDF-1 and H7, a developmentally regulated gene isolated in D. discoideum (18,19) (Fig. 1C).…”

Section: Edf-1 Expression In Differentiating Endothelialmentioning

confidence: 99%

EDF-1, a Novel Gene Product Down-regulated in Human Endothelial Cell Differentiation

Dragoni¹,

Mariotti²,

Consalez³

et al. 1998

Journal of Biological Chemistry

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“…1) consisting of the Dictyostelium V18 promoter, a polylinker, and the terminator of actin 8, on a vector containing a neomycin-resistance cassette. The promoter from the V18 gene has been shown to be active in vegetative cells and in starving cells up to 8-10 hours after the onset of starvation (20,23). The antisense vector was designed so that transformed cells will become resistant to neomycin and express antisense RNA complementary to the mRNA from which the cDNA was derived.…”

Section: Resultsmentioning

confidence: 99%

Mutagenesis and gene identification in Dictyostelium by shotgun antisense.

Spann

Brock

Lindsey

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

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We have developed a mutagenesis technique that uses antisense cDNA to identify genes required for development in Dictyostelium discoideum. We and 2).There is no general way to map or rescue chemically induced mutations. However, both extrachromosomal and integrating transformation vectors exist for Dictyostelium (3-8), and gene expression can be repressed by both antisense RNA and homologous recombination-mediated gene disruption (9-16). Mutants can also be generated by random insertion of transformation plasmids into the genome (17,18). A portion of the disrupted gene can then be isolated by digesting the DNA of the mutant and recircularizing the DNA so that the resulting plasmid contains genomic DNA flanking the insertion site. In the Kuspa and Loomis technique (restriction enzyme-mediated insertion; REMI), the excised plasmid can then be relinearized and used for homologous recombination to verify that disruption of the gene has caused the mutant phenotype. A limitation of knock-out strategies is that the disruption of many genes is lethal to the cell.In contrast, antisense repression can permit a substantial reduction but not a complete block in the synthesis of the corresponding gene products. This has allowed the investigation of the effect of decreased amounts of proteins such as calmodulin, where complete repression is lethal (19). Another advantage of antisense repression is that transformation with a single construct will repress expression of proteins from multiple genes that express essentially identical transcripts, such as the three-member Discoidin I gene family (9). Similar repression by homologous recombination would require three separate rounds of transformation. In this report, we describe a mutagenesis scheme for Dictyostelium based on antisense transformation. This method allows the sequence of the repressed mRNA to be examined a few days after isolation of an interesting phenotype and also allows the identification of genes that would not be detected in other mutagenesis strategies. MATERIALS AND METHODSConstruction of pV18neo Antisense Vector. The Dictyostelium actin 8 terminator contained in the HindIlI-EcoRI fragment of pDneoll (12) was ligated into HindIII/EcoRI-digested pBluescript SKI (Stratagene). After digestion with PstI and HindIlI, the fragment containing the terminator was ligated into PstI/HindIII-digested pSP72. The XbaI site of the resulting plasmid was destroyed by digesting with BamHI and Sall and ligation with a mix of the oligos GATCCGCCCGCGACG and TCGACGTCGCGGGCG to create pSP72A8, preserving the BamHI and Sall sites into which the cDNA library was later ligated. The V18 promotor fragment was isolated as a BamHI/ HindIII fragment from pVl8p3-90 (20) and was ligated into the BamHI and HindIII sites of pBluescript SKI. A ClaI/BamHI fragment containing the promotor was isolated from the resulting plasmid and was ligated with the 3-kb fragment from a BamHI/ partial ClaI digest of pSP72A8 to produce pHC. The BamHI and SalI sites of pUC 19 were destroyed by digesti...

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“…Approximately 25 % of the estimated 8000 genes in the Dictyostelium genome are regulated during the transition from unicellular to multicellular development [10]. So far, a large number of genes that are temporally regulated during development have been reported, and most of them participate in the progression of development [1,2,5,11]. However, the mechanism of transcriptional regulation of these genes remains to be solved.…”

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confidence: 99%

DNG1, a Dictyostelium homologue of tumor suppressor ING1 regulates differentiation of Dictyostelium cells

Mayanagi

Amagai

Maeda

2005

CMLS, Cell. Mol. Life Sci.

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dng1 is a Dictyostelium homologue of the mammalian tumor suppressor ING gene. DNG1 protein localizes in the nucleus, and has a highly conserved PHD finger domain found in chromatin-remodeling proteins. Both dng1 disruption and overexpression impaired cell proliferation. In dng1-null cells, the progression of differentiation was delayed in a cell-density-dependent manner, and many tiny aggregates were formed. Exogenously applied cAMP pulses reversed the inhibitory effect caused by dng1 disruption on the aggregation during early development, but formation of tiny aggregates was not restored. dng1-overexpressing cells acquired the ability to undergo chemotaxis to cAMP earlier and exhibited enhanced differentiation. These phenotypes were found to be coupled with altered expressions of early genes such as cAMP receptor 1 (car1) and contact site A (csA). Furthermore, disordered histone modifications were demonstrated in dng1-null cells. These results suggest a regulatory role of dng1 in the transition of cells from growth to differentiation.

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Primary structure and regulation of vegetative specific genes ofDictyostelium discoideum

Cited by 47 publications

References 28 publications

EDF-1, a Novel Gene Product Down-regulated in Human Endothelial Cell Differentiation

EDF-1, a Novel Gene Product Down-regulated in Human Endothelial Cell Differentiation

Mutagenesis and gene identification in Dictyostelium by shotgun antisense.

DNG1, a Dictyostelium homologue of tumor suppressor ING1 regulates differentiation of Dictyostelium cells

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