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Volumn 277, Issue 5323, 1997, Pages 251-254

Dictyostelium development in the absence of cAMP

Author keywords

[No Author keywords available]

Indexed keywords

CYCLIC AMP; CYCLIC AMP DEPENDENT PROTEIN KINASE;

EID: 0030873976     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5323.251     Document Type: Article
Times cited : (90)

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    • Franke, J.1    Podgorski, G.J.2    Kessin, R.H.3
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    • note
    • To determine the extent of sporulation, we transferred sori from individual fruiting bodies to 20 μl of 20 mM potassium phosphate buffer (pH 6.2) containing 0.4% NP-40 nonionic detergent (Sigma) and triturated at room temperature. The number of refractile, ovoid spores was determined by phase-contrast microscopy using a hemocytometer. Spores were counted for five fruiting bodies produced by wild-type or acaA(PKA-C) cells.
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    • note
    • The viability of the spores was determined after detergent treatment (25) by plating 1500 visible spores on 10 SM agar plates in association with Klebsiella aerogenes (27). The number of colonies formed was assumed to be representative of the number of viable spores in the original sample.
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    • note
    • 2 circle was formed after the buffer was absorbed by the agar. Total numbers of aggregates and fruiting bodies formed were scored on a dissecting microscope after 14 hours and 36 hours of development, respectively. At the highest cell density tested, some of the aggregates formed by acaA(PKA-C) cells, which did not progress to normal fruiting bodies, formed terminal structures similar to those observed for wild-type cells overexpressing pkaC (7, 8) or pkaR-null mutants (6). The acaA mutant cells were never observed to produce aggregates at these cell densities (23). A similar cell density dependence for aggregation of the acaA(PKA-C) cells was found on Millipore filters (23).
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    • note
    • We thank W. F. Loomis, D. Hereld, and S. Lu for many helpful discussions regarding this work; G. Souza, G. Shaulsky, and W. F. Loomis for suggestions that improved the manuscript; and R. Sucgang for help in preparing the figures. Supported by grants from NIH and the State of Texas. A.K. is a Searle Scholar and an American Cancer Society Junior Faculty Research Fellow.


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