Null mutations of the Dictyostelium cyclic nucleotide phosphodiesterase gene block chemotactic cell movement in developing aggregates.
Extracellular cAMP is a critical messenger in the multicellular development of the cellular slime mold Dictyostelium discoideum. The levels of cAMP are controlled by a cyclic nucleotide phosphodiesterase (PDE) that is secreted by the cells. The PDE gene (pdsA) is controlled by three promoters that permit expression during vegetative growth, during aggregation, and in prestalk cells of the older structures. Targeted disruption of the gene aborts development, and complementation with a modified pdsA restores development. Two distinct promoters must be used for full complementation, and an inhibitory domain of the PDE must be removed. We took advantage of newly isolated PDE-null cells and the natural chimerism of the organism to ask whether the absence of PDE affected individual cell behavior. PDE-null cells aggregated with isogenic wild-type cells in chimeric mixtures, but could not move in a coordinated manner in mounds. The wild-type cells move inward toward the center of the mound, leaving many of the PDE-null cells at the periphery of the aggregate. During the later stages of development, PDE-null cells in the chimera segregate to regions which correspond to the prestalk region and the rear of the slug. Participation in the prespore/spore population returns with the restoration of a modified pdsA to the null cells.
Sucgang
R
R
College of Physicians and Surgeons, Columbia University, 630 West 168 Street, New York, New York 10032, USA.
Weijer
C J
CJ
Siegert
F
F
Franke
J
J
Kessin
R H
RH
D015105:3',5'-Cyclic-AMP Phosphodiesterases / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D002633:Chemotaxis / Q000235:genetics; D002678:Chimera; D017931:DNA Primers / Q000235:genetics; D004023:Dictyostelium / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development*; D018390:Gene Targeting; D005800:Genes, Fungal; D017125:Genes, Protozoan; D005816:Genetic Complementation Test; D009154:Mutation; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion
D017931:DNA Primers; D015105:3',5'-Cyclic-AMP Phosphodiesterases