Cells respond to and bind countin, a component of a multisubunit cell number counting factor.
In Dictyostelium discoideum counting factor (CF), a secreted approximately 450-kDa complex of polypeptides, inhibits group and fruiting body size. When the gene encoding countin (a component of CF) was disrupted, cells formed large groups. We find that recombinant countin causes developing cells to form small groups, with an EC(50) of approximately 3 ng/ml, and affects cAMP signal transduction in the same manner as semipurified CF. Recombinant countin increases cell motility, decreases cell-cell adhesion, and regulates gene expression in a manner similar to the effect of CF. However, countin does not decrease adhesion or group size to the extent that semipurified CF does. A 1-min exposure of developing cells to countin causes an increase in F-actin polymerization and myosin phosphorylation and a decrease in myosin polymerization, suggesting that countin activates a rapid signal transduction pathway. (125)I-Labeled countin has countin bioactivity, and binding experiments suggest that vegetative and developing cells have approximately 53 cell-surface sites that bind countin with a K(D) of approximately 1.5 ng/ml or 60 pm. We hypothesize that countin regulates cell development through the same pathway as CF and that other proteins within the complex may modify the activity of countin and/or have independent size-regulating activities.
Gao
Tong
T
Howard Hughes Medical Institute, Rice University, Houston, Texas 77005-1892, USA.
Ehrenman
Karen
K
Tang
Lei
L
Leippe
Matthias
M
Brock
Debra A
DA
Gomer
Richard H
RH
D000199:Actins / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D002465:Cell Movement; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D004023:Dictyostelium / Q000378:metabolism*; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D007455:Iodine / Q000494:pharmacology; D007700:Kinetics; D009218:Myosins / Q000378:metabolism; D010766:Phosphorylation; D011485:Protein Binding; D015800:Protozoan Proteins / Q000737:chemistry* / Q000502:physiology*; D011994:Recombinant Proteins / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D014461:Ultracentrifugation
D000199:Actins; D018076:DNA, Complementary; D015800:Protozoan Proteins; D011994:Recombinant Proteins; C120918:countin protein, Dictyostelium discoideum; D007455:Iodine; D000242:Cyclic AMP; D009218:Myosins