Separation and properties of prestalk and prespore cells of Dictyostelium discoideum

Tsang, Adrian; Bradbury, Jane M.

doi:10.1016/0014-4827(81)90118-x

Cited by 81 publications

(33 citation statements)

References 27 publications

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“…The anterior one-fifth of the migratory slug of Dictyostelium discoideum contains stalk cell precursors, while the posterior four-fifths consists predominantly of prespore cells. Genes, which are expressed in a cell-typespecific manner during development, are readily isolated (1,17,27), because prestalk-and prespore-cell-enriched populations can be separated by microdissection or by density gradient centrifugation of disaggregated slugs (33,42). The D19 cDNA clone derives from a highly prespore-enriched mRNA obtained from density gradient-separated cells (1).…”

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confidence: 99%

Structural characterization of Dictyostelium discoideum prespore-specific gene D19 and of its product, cell surface glycoprotein PsA.

Early¹,

Williams²,

Meyer³

et al. 1988

Mol. Cell. Biol.

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The Dictyostelium discoideum cell surface antigen PsA is a glycoprotein which first appears in the multicellular stage soon after tip formation and is selectively expressed on prespore cells. The D19 gene encodes an mRNA sequence which is highly enriched in prespore over prestalk cells in the slug stage. We have determined 81 amino acid residues of N-terminal sequence from immunoaffinity-purified PsA protein and shown this sequence to be identical to the predicted sequence of the D19 gene. There are several short repeat elements close to the C terminus, and unequal crossing-over within these is proposed to account for the size polymorphism observed in PsA protein isolated from different D. discoideum strains. The repeats are proline rich and show similarity to the C-terminal region of the D. discoideum cell adhesion molecule, contact sites A. The extreme C terminus, which is also homologous to contact sites A, is characteristic of proteins attached to the plasma membrane via a glycosyl-phosphatidylinositol link. We have marked the PsA gene by insertion of an oligonucleotide encoding an epitope of the human c-myc protein. A construct containing this gene and 990 base pairs of 5'-flanking region directed correct temporal and spatial mRNA accumulation. We found the marked PsA protein, detected with the human c-myc antibody, to be correctly localized on the surface of cells.

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mentioning

confidence: 99%

Structural characterization of Dictyostelium discoideum prespore-specific gene D19 and of its product, cell surface glycoprotein PsA.

Early¹,

Williams²,

Meyer³

et al. 1988

Mol. Cell. Biol.

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“…Addition of cAMP to the suspension of cells dispersed from the aggregates restores transcription (13) and prevents destabilization (6). This class of mRNAs appears to be preferentially expressed in pre-spore cells (18,19). In contrast, dispersion of cell aggregates does not affect the synthesis and stability of the constitutive mRNAs and of a second, smaller class of developmentally regulated mRNA species that appears to be preferentially expressed in pre-stalk cells (refs.…”

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confidence: 99%

mRNA stabilization controls the expression of a class of developmentally regulated genes in Dictyostelium discoideum

Mangiarotti

Giorda

Ceccarelli

et al. 1985

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

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During the development of Dictyostelium discoideum, several thousand new mRNA species appear in the cytoplasm after the cells have formed stable aggregates. Here we show that six of these late mRNAs, corresponding to six clones randomly chosen from a genomic library, are synthesized from the very beginning of development at a rate comparable to that observed late in development but that transcripts do not accumulate until after aggregation. The early-and late-synthesized mRNAs are identical in size and compete with each other for hybridization to the genomic clones. The early-synthesized mRNAs do not accumulate in the cytoplasm in the preaggregation stage because they are very unstable. Their stability, estimated from the kinetics of incorporation during continuous labeling with 32p, increases by perhaps an order of magnitude in the postaggregation stage. We conclude that mRNA stabilization is the major controlling factor of the expression of these genes.

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“…We observed a second burst of synthesis of cAMP-dependent protein kinase at culmination (11); therefore, it is quite possible that the cAMP-dependent protein kinase plays a role in the initiation of culmination. Tsang and Bradbury (20) and Ratner and Borth (24) …”

Section: Discussionmentioning

confidence: 99%

“…Amoebae were allowed to develop until they reached the slug stage. The procedure for the separation was a modification of that described by Tsang and Bradbury (20). The aggregates were disrupted in 50 mM Tris, pH 7.0/13 mM dimercaptopropanol/0.01% Pronase (BAL/Pronase).…”

Section: Methodsmentioning

confidence: 99%

Differential cellular distribution of cAMP-dependent protein kinase during development of Dictyostelium discoideum

Schaller

Leichtling

Majerfeld

et al. 1984

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

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It was shown previously by us that cAMPdependent protein kinase activity in the cellular slime mold Dictyostelium discoideum increased during the early stages of development. Results from other laboratories showed that during the subsequent stage of cell differentiation and positioning, the accumulation of a number of prespore mRNAs and proteins (but not prestalk mRNAs and proteins) was dependent upon cAMP. The present communication describes the cellular distribution of the cAMP-dependent protein kinase at that stage of development. Pseudoplasmodia were disrupted, and prespore cells were separated from prestalk cells by sedimentation through a Percoll gradient. Prespore cells had approximately 4-5 times as much of both the catalytic and regulatory subunits of the cAMP-dependent protein kinase as did the prestalk cells. That the increase of cAMP-dependent protein kinase during development reflected de novo synthesis of the enzyme in both prespore and prestalk cells was demonstrated on the basis of [3H]leucine incorporation into the regulatory subunit. The findings are consistent with a role of the cAMP-dependent protein kinase in mediating the effects of cAMP on the synthesis of prespore-specific mRNAs and proteins at the stage at which cAMP appears to be required for the cell type-specific syntheses.The cellular slime mold Dictyostelium discoideum is a favorite organism for the study of cellular differentiation and morphogenesis. Cyclic AMP plays a central role in the development of the organism. The amoebae, when deprived of nutrients, eventually give rise to fruiting bodies consisting of three differentiated types of cells-spores supported by a column of nonviable stalk cells resting on a basal disk also composed of nonviable cells.Cellular differentiation is apparent once the pseudoplasmodium (slug) has been formed; amoebae in the anterior fourth of the slug eventually will become stalk cells, while the amoebae in the posterior portion are predestined to become primarily spores; a small population of "anterior-like" cells, which are associated with the posterior part of the slug, will form the basal disk during culmination. The anterior and posterior cells can be distinguished by morphological and biochemical criteria including the occurrence of specific mRNAs, proteins, and subcellular organelles (1).The implementation of the first stage of the developmental program from the onset of starvation to the formation of loose aggregates does not require cell-cell contact and is stimulated by exogenous cAMP (2). This stage of development is characterized by the appearance of new mRNAs and proteins, which are formed presumably in all amoebae up to aggregation. During aggregation (prior to tip formation), another class of mRNAs and proteins appears, which also probably is found in all aggregating amoebae; but, at a later stage of development, its occurrence is confined to the anterior part of the slug (3-5). A third family of mRNAs and proteins first appears concomitant with the formation of tips on the aggr...

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Separation and properties of prestalk and prespore cells of Dictyostelium discoideum

Cited by 81 publications

References 27 publications

Structural characterization of Dictyostelium discoideum prespore-specific gene D19 and of its product, cell surface glycoprotein PsA.

Structural characterization of Dictyostelium discoideum prespore-specific gene D19 and of its product, cell surface glycoprotein PsA.

mRNA stabilization controls the expression of a class of developmentally regulated genes in Dictyostelium discoideum

Differential cellular distribution of cAMP-dependent protein kinase during development of Dictyostelium discoideum

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