The timing of phenotypic suppression of an aggregation defect by an aggregation-stimulating factor from Polyspboadylium violaceum

Hanna, Michael H.; Gardner, Jeffrey L.; Nowicki, J J

doi:10.1111/j.1432-0436.1984.tb01343.x

Differentiation

1984

DOI: 10.1111/j.1432-0436.1984.tb01343.x

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The timing of phenotypic suppression of an aggregation defect by an aggregation-stimulating factor from Polyspboadylium violaceum

Michael H. Hanna

Jeffrey L. Gardner

J J Nowicki

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1984

1988

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Cited by 5 publications

(7 citation statements)

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“…One possible explanation is that the aggA mutation is not in the structural gene for the enzyme responsible for D-factor biosynthesis but that the aggA function is involved in the same developmental pathway as D factor. While we cannot rule out this possibility, it seems unlikely, since all non-aggA Agg-mutants previously isolated produce D factor (4; Hanna, unpublished data), and D factor is known to affect a late preaggregation function (4,6). This explanation does not address the fact that other biosynthetic mutants were not found.…”

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confidence: 43%

“…The relative proportions of the individual components are developmentally and environmentally regulated, with light having a significant effect on the total production of D factor and the proportions of the individual components (1,5). The pheromone D factor is most likely involved in the chemotactic process (9) and not in cellular adhesion (6). D factor is required only just prior to aggregation, possibly triggering autonomous production of glorin or development of founder cells (4,6,9,13).…”

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“…The pheromone D factor is most likely involved in the chemotactic process (9) and not in cellular adhesion (6). D factor is required only just prior to aggregation, possibly triggering autonomous production of glorin or development of founder cells (4,6,9,13). Founder cells are the first cells to produce glorin and are morphologically distinct in P. violaceum (10,13).…”

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Analysis of developmentally defective chemical signaling mutants of Polysphondylium violaceum

Ellsaesser¹,

Kühn²,

Hanna³

1986

J Bacteriol

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Six aggregation-defective mutants of Polysphondylium violaceum dependent on external addition of the pheromone D factor for aggregation were isolated after nitrosoguanidine mutagenesis. With a screening technique based on synergistic development, D-factor-dependent mutants can be separated from other kinds of aggregateless mutants. Genetic complementation analyses of the newly isolated mutants showed them to be mutant at the aggA locus. Individual mutants exhibited different sensitivities to D factor(s), responding maximally over a 300-fold range of concentrations.

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confidence: 43%

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confidence: 99%

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confidence: 99%

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Analysis of developmentally defective chemical signaling mutants of Polysphondylium violaceum

Ellsaesser¹,

Kühn²,

Hanna³

1986

J Bacteriol

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“…Since these components of the aggregation system have developed in the aggA mutants, D factor is suggested to be a late-acting component of the aggregation system. Further evidence (10) that D factor affects a late aggregation function was shown in experiments where aggA mutants starved without D factor for several hours would aggregate within 15-30 min after the addition of D factor; whereas, if D factor was added to these cells at the beginning of starvation, aggregation occurred after 5 hr.…”

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confidence: 96%

“…D factor has been shown to be a family of related compounds (9) and one member of this family, Df,, has been purified (FATONE et al, submitted). Although unable to aggregate in the absence of D factor, aggA mutants make stable cell-cell contacts characteristic of aggregation-competent wild-type amoebae (10). In addition, these mutants exhibit normal chemotactic sensitivity and response without added D factor (11).…”

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confidence: 99%

Founder Cell Differentiation and Acrasin Production in an Aggregateless Mutant of Polyspondylium violaceum

Newth¹,

Arnal²,

Goulart³

et al. 1987

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A specific class of aggregation-deficient mutants, aggA, of Polysphondyliurn violaceurn are unable to aggregate unless supplied exogenously with a stimulating factor called D factor. The present study examines the effect of D factor on the induction of founder cells and on the production of the chemoattractant of aggregation, N-propionyl--L-glutamyl-L-ornithine-6 -1actam ethyl ester (or glorin). Founder cells initiate aggregate formation and are morphologically distinct from the majority of the amoebae. Founder cell differentiation and oriented movement of attracted amoebae have been studied by time-lapse videotape analysis. In wild-type strains, on the average 90 min after the onset of starvation, a single, motile, irregularly shaped amoeba stops wandering and becomes round in shape. This founder cell has differentiated randomly from the pool of starved amoebae and within 2.5 min after the cessation of movement begins to attract and establish cellular contacts with neighboring amoebae. The aggA mutants neither aggregate nor differentiate founder cells in the absence of D factor; whereas, aggregate formation and founder cell differentiation occur in the presence of physiological concentrations of purified, externally added D factor. However, in either the presence or absence of D factor, aggA amoebae produce and excrete glorin (measured using a bioassay) at levels comparable to their parental strain. These studies suggest that D factor is required for founder cell differentiation and organization of the aggregate, and that the ability to synthesize and excrete glorin is not sufficient to trigger aggregation.In Polysphondylium violaceum, aggregation is inititated by a single morphologically identifiable cell (1). This amoeba is called a founder cell and is round and non-motile in contrast to the majority of the amoebae which are irregularly shaped and motile. Founder cell are not a predetermined class of cells, but arise from the general population of amoebae just prior to aggregate formation (1,2, and this paper). Founder cells are believed to initiate aggregate formation by chemotactically attracting wandering amoebae to a stationary center. The chemoattractant for aggregation in P. violaceum is a modified peptide called glorin (N-propionyl-Y -L-glutamyl-L-ornithine-6 -1actam ethyl ester, ref. 3) and not cyclic AMP. In Dictyostelium morphologically distinct founder cells have not been observed. Aggregation in D. discoideum begins when a random amoeba or group of amoebae begin release of the chemoattractant 3', 5' cyclic adenosine monophosphate (cyclic AMP) in response to starvation conditions (4). Both P. violaceum and D. discoideum are likely to initiate aggregate formation through autonomous pulsatile release of their specific chemoattractant.

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Purification, characterization, and partial structure of D factor from Polysphondylium violaceum

et al. 1988

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The A component of D factor (DfA) was overproduced during development of wild type Polyspondylium violaceum strain China after starvation in liquid medium. Crude DfA excreted by strain China was partially purified by ultrafiltration using Amicon YM10 and YM2 filters with DfA extracted from the filtrate by absorption onto a preparative grade C-18 resin. The concentrated material was further purified on a C-18 analytical column using both acetonitrile:water and methanol:water gradients. This highly purified fraction was a single component with a final specific activity of greater than 10(6) units per mg dry weight. Purified DfA is red having a broad visible absorbance at 500 nm and a ultraviolet (uv) absorbance at 290-300 nm. The red chromophore is sensitive to pH and to oxidation-reduction. 1H and 13C nmr studies with purified DfA indicate that it is a C11 compound with both polar and non-polar regions. The non-polar region has been identified as a hexanone and is the same as the side chain of DIF from Dictyostelium discoideum. Purified DfA has been used in studies with the D factor non-producing mutant, tsg-119 cyc-1 aggA586 (A586), to show that neither production of glorin nor chemotactic sensitivity to glorin are affected by D factor. However, founder cells develop in A586 mutant populations only after addition of D factor. These data suggest that DfA may be necessary for induction of aggregate formation by aggregation-competent amoebae.

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The timing of phenotypic suppression of an aggregation defect by an aggregation-stimulating factor from Polyspboadylium violaceum

Cited by 5 publications

References 14 publications

Analysis of developmentally defective chemical signaling mutants of Polysphondylium violaceum

Analysis of developmentally defective chemical signaling mutants of Polysphondylium violaceum

Founder Cell Differentiation and Acrasin Production in an Aggregateless Mutant of Polyspondylium violaceum

Purification, characterization, and partial structure of D factor from Polysphondylium violaceum

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