Selection of chemotaxis mutants of Dictyostelium discoideum

J. E. Segall, P. R. Fisher, G. Gerisch

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

A method has been developed for the efficient selection of chemotaxis mutants of Dictyostelium discoideum. Mutants defective in the chemotactic response to folate could be enriched up to 30-fold in one round of selection using a chamber in which a compartment that contained the chemoattractant was separated by a sandwich of four nitrocellulose filters from a compartment that contained buffer. Mutagenized cells were placed in the center of the filter layer and exposed to the attactant gradient built up between the compartments for a period of 3-4 h. While wild-type cells moved through the filters in a wave towards the compartment that contained attractant, mutant cells remained in the filter to which they were applied. After several repetitions of the selection procedure, mutants defective in chemotaxis made up 10% of the total cell population retained in that filter. Mutants exhibiting three types of alterations were collected: (a) motility mutants with either reduced speed of movement, or altered rates of turning; (b) a single mutant defective in production of the attractant-degrading enzyme, folate deaminase; and (c) mutants with normal motility but reduced chemotactic responsiveness. One mutant showed drastically reduced sensitivity in folate-induced cGMP production. Morphogenetic alterations of mutants defective in folate chemotaxis are described.

Original languageEnglish (US)
Pages (from-to)151-161
Number of pages11
JournalJournal of Cell Biology
Volume104
Issue number1
DOIs
StatePublished - 1987

ASJC Scopus subject areas

  • Cell Biology

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