Identification of an endogenous plasmid in <i>Dictyostelium discoideum</i>

Metz, Birgit A.; Ward, T.E.; Welker, Dennis L.; Williams, Keith L.

doi:10.1002/j.1460-2075.1983.tb01456.x

Cited by 60 publications

(31 citation statements)

References 26 publications

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“…220 I I expression of some genes is limited to particular times of development. Such studies should be facilitated by construction of cloning vectors for D. discoideum and this may be possible from a recently discovered D. discoideum plasmid (Metz et al, 1983).…”

Section: Discussionmentioning

confidence: 99%

Protein synthesis during development and differentiation in the cellular slime mould Dictyostelium discoideum

Watts¹

1984

Biochemical Journal

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Section: Discussionmentioning

confidence: 99%

Protein synthesis during development and differentiation in the cellular slime mould Dictyostelium discoideum

Watts¹

1984

Biochemical Journal

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“…Diploids were constructed as previously described (Newell et al, 1977), except that selection of the diploid DU2413 was based on complementation of the recessive bsgA.5 mutation and the dominance of the cob354 cobalt-resistance mutation (Williams, 1978;Metz et al, 1983). For this selection, the cobalt-resistant Bacillus subtilis strain BSU2, isolated from strain 36.1 by Dr S. Kayman in our laboratory, was incubated overnight on SM agar containing 250 pg cobaltous chloride ml-and 10 pg dihydrostreptomycin sulphate ml-', to form a thin lawn prior to inoculation with D. discoideum.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, to select diploids we used a haploid tester strain, HU 1628, which carried the unstable dominant cobalt resistance cob-354 (Williams, 1978;Metz et al, 1983) and also markers on all six established linkage groups (Table 1). The resulting diploid (DU2413) was heterozygous with respect to CAMPS resistance, as well as to the markers of the tester strain (Table 1).…”

Section: Assignment Of a Camps-resistance Mutation To Linkage Group IImentioning

confidence: 99%

Genetic Analysis of a Dictyostelium discoideum Mutant Resistant to Adenosine 3':5'-Cyclic Phosphorothioate, an Inhibitor of Wild-type Development

Wallraff¹,

Welker²,

Williams³

et al. 1984

Microbiology

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A mutant of Dictyostelium discoideurn strain AX2 with altered responses to cyclic AMP has been analysed genetically. The mutant, HG302, aggregates and fruits in the presence of adenosine 3' : 5'-cyclic phosphorothioate (CAMPS). This slowly hydrolysed analogue of cyclic AMP inhibits wild-type development by blocking cell differentiation from the growth phase to the aggregation-competent stage. The mutant forms small aggregates in the absence and in the presence of CAMPS, and is further distinguished from the wild type by decreased activities of extracellular phosphodiesterase and phosphodiesterase inhibitor. These phenotypic characteristics were all linked to the CAMPS resistance, which was assigned to linkage group 11. Mitotic crossover experiments showed that the CAMPS-resistance mutation, casA1000, maps proximal to the acrA locus in the neighbourhood of whiA. The data suggest that casAlOO0 acts pleiotropically, and that the product of the wild-type allele is required in a part of the signalprocessing pathway common to control of development and regulation of extracellular phosphodiesterase.

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“…All are found in multiple copies, and their sizes vary from approximately 7 to 25 kilobases (kb) (11; K. Williams personal communication; D. Welker, personal communication; R. A. Firtel and C. Reymond, unpublished data). The three plasmids that have been analyzed to date do not appear to cross-hybridize under standard conditons, suggesting that they do not share any major nucleotide sequence homology (R. A. Firtel, unpublished data).…”

mentioning

confidence: 99%

Extrachromosomal replication of shuttle vectors in Dictyostelium discoideum.

Firtel¹,

Silan²,

Ward³

et al. 1985

Mol. Cell. Biol.

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We cloned a 12.3-kilobase (kb) endogenous plasmid, Ddpl, found in several wild-type and laboratory strains of Dictyostelium discoideum into pBR322. The cloned plasmids have been used to cotransform D. discoideum cells with B1OS, a transformation vector carrying a gene fusion conferring resistance to G418. Whereas BIOS DNA alone appears to integrate in a tandem array, the cloned Ddpl plasmids replicate extrachromosomally and are stably maintained in the absence of selection with an average copy number of 50 to 100 copies per cell. The Ddpl-derived plasmids can be directly recovered by transforming Escherichia coli with bulk nuclear DNA from these cells. Preliminary deletion analysis indicates that not all regions of Ddpl are necessary for stable replication in D. discoideum. Several recombinant vectors which replicate extrachromosomally in D. discoideum were also isolated. One contains the Act6-neor gene fusion from BiOS recombined into one of the cloned derivatives of Ddpl and can be used to directly transform D. discoideum amoebae, selecting for G418 resistance. Another recombinant is only 5.6 kb and resulted from a deletion of a 16.6-kb cloned Ddpl hybrid plasmid. An analysis of the vector DNAs present in clones derived from single D. discoideum transformants is also described.Dictyosteliium discoideium grows as single-celled amoebae. Upon starvation, the cells initiate a multicellular developmental program in which approximately 105 cells form an aggregate which then differentiates into a fruiting body containing predominantly two cell types: spores and stalk cells (7). We and others have been interested in understanding the mechanisms by which cell-type differentiation is regulated in this relatively simple multicellular system. To this end, genes which are expressed at specific developmental stages were cloned and used to examine the biochemical and physiological factors which regulate their expression. These genes include those expressed during vegetative growth and repressed during early development, genes expressed under various conditions during preaggregation, and the cell-type-specific genes that are preferentially expressed in either prestalk or prespore cells (1, 9; for a review, see reference 4). Several factors have been identified which are required for the expression of sets of coordinately regulated genes, including cAMP (3, 5, 8-10; S. Mann, R. A. Firtel, and J. Brandis, manuscript in preparation).We have begun by examining the cis-acting DNA sequences that are involved in the proper developmental regulation of the expression of these genes and have established that there is a DNA-mediated transformation system in which cloned genes can be transformed into stably maintained and expressed D. discoidelum (12, 13 Firtel; Proc. Natl. Acad. Sci. USA, in press; W. Nellen and R. A. Firtel, Gene, in press). For several types of analyses, it would be advantageous to have a shuttle vector which would replicate extrachromosomally in D. discoideutm cells and which could be easily recovered by transforming...

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Identification of an endogenous plasmid in Dictyostelium discoideum

Cited by 60 publications

References 26 publications

Protein synthesis during development and differentiation in the cellular slime mould Dictyostelium discoideum

Protein synthesis during development and differentiation in the cellular slime mould Dictyostelium discoideum

Genetic Analysis of a Dictyostelium discoideum Mutant Resistant to Adenosine 3':5'-Cyclic Phosphorothioate, an Inhibitor of Wild-type Development

Extrachromosomal replication of shuttle vectors in Dictyostelium discoideum.

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