Chemotaxis to cAMP and slug migration in Dictyostelium both depend on migA, a BTB protein.
Chemotaxis in natural aggregation territories and in a chamber with an imposed gradient of cyclic AMP (cAMP) was found to be defective in a mutant strain of Dictyostelium discoideum that forms slugs unable to migrate. This strain was selected from a population of cells mutagenized by random insertion of plasmids facilitated by introduction of restriction enzyme (a method termed restriction enzyme-mediated integration). We picked this strain because it formed small misshapen fruiting bodies. After isolation of portions of the gene as regions flanking the inserted plasmid, we were able to regenerate the original genetic defect in a fresh host and show that it is responsible for the developmental defects. Transformation of this recapitulated mutant strain with a construct carrying the full-length migA gene and its upstream regulatory region rescued the defects. The sequence of the full-length gene revealed that it encodes a novel protein with a BTB domain near the N terminus that may be involved in protein-protein interactions. The migA gene is expressed at low levels in all cells during aggregation and then appears to be restricted to prestalk cells as a consequence of rapid turnover in prespore cells. Although migA- cells have a dramatically reduced chemotactic index to cAMP and an abnormal pattern of aggregation in natural waves of cAMP, they are completely normal in size, shape, and ability to translocate in the absence of any chemotactic signal. They respond behaviorally to the rapid addition of high levels of cAMP in a manner indicative of intact circuitry connecting receptor occupancy to restructuring of the cytoskeleton. Actin polymerization in response to cAMP is also normal in the mutant cells. The defects at both the aggregation and slug stage are cell autonomous. The MigA protein therefore is necessary for efficiently assessing chemical gradients, and its absence results in defective chemotaxis and slug migration.
Escalante
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R
Department of Biology, University of California San Diego, La Jolla 92093, USA.
Wessels
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D
Soll
D R
DR
Loomis
W F
WF
D000595:Amino Acid Sequence; D000818:Animals; D015415:Biomarkers; D002449:Cell Aggregation; D002633:Chemotaxis / Q000187:drug effects / Q000235:genetics; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000494:pharmacology*; D004023:Dictyostelium / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D017353:Gene Deletion; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D010641:Phenotype; D015800:Protozoan Proteins
D015415:Biomarkers; D005656:Fungal Proteins; C109071:MigA protein, Dictyostelium; D015800:Protozoan Proteins; D000242:Cyclic AMP