TY - JOUR
T1 - Two Components of a Secreted Cell Number-counting Factor Bind to Cells and Have Opposing Effects on cAMP Signal Transduction in Dictyostelium
AU - Brock, Debra A.
AU - Ehrenman, Karen
AU - Ammann, Robin
AU - Tang, Yitai
AU - Gomer, Richard H.
PY - 2003/12/26
Y1 - 2003/12/26
N2 - A secreted 450-kDa complex of proteins called counting factor (CF) is part of a negative feedback loop that regulates the size of the groups formed by developing Dictyostelium cells. Two components of CF are countin and CF50. Both recombinant countin and recombinant CF50 decrease group size in Dictyostelium. countin-cells have a decreased cAMP-stimulated cAMP pulse, whereas recombinant countin potentiates the cAMP pulse. We find that cf50- cells have an increased cAMP pulse, whereas recombinant CF50 decreases the cAMP pulse, suggesting that countin and CF50 have opposite effects on cAMP signal transduction. In addition, countin and CF50 have opposite effects on cAMP-stimulated Erk2 activation. However, like recombinant countin, recombinant CF50 increases cell motility. We previously found that cells bind recombinant countin with a Hill coefficient of ∼2, a KH of 60 pM, and ∼53 sites/cell. We find here that cells also bind 125I-recombinant CF50, with a Hill coefficient of ∼2, a K H of ∼15 ng/ml (490 pM), and ∼56 sites/cell. Countin and CF50 require each other's presence to affect group size, but the presence of countin is not necessary for CF50 to bind to cells, and CF50 is not necessary for countin to bind to cells. Our working hypothesis is that a signal transduction pathway activated by countin binding to cells modulates a signal transduction pathway activated by CF50 binding to cells and vice versa and that these two pathways can be distinguished by their effects on cAMP signal transduction.
AB - A secreted 450-kDa complex of proteins called counting factor (CF) is part of a negative feedback loop that regulates the size of the groups formed by developing Dictyostelium cells. Two components of CF are countin and CF50. Both recombinant countin and recombinant CF50 decrease group size in Dictyostelium. countin-cells have a decreased cAMP-stimulated cAMP pulse, whereas recombinant countin potentiates the cAMP pulse. We find that cf50- cells have an increased cAMP pulse, whereas recombinant CF50 decreases the cAMP pulse, suggesting that countin and CF50 have opposite effects on cAMP signal transduction. In addition, countin and CF50 have opposite effects on cAMP-stimulated Erk2 activation. However, like recombinant countin, recombinant CF50 increases cell motility. We previously found that cells bind recombinant countin with a Hill coefficient of ∼2, a KH of 60 pM, and ∼53 sites/cell. We find here that cells also bind 125I-recombinant CF50, with a Hill coefficient of ∼2, a K H of ∼15 ng/ml (490 pM), and ∼56 sites/cell. Countin and CF50 require each other's presence to affect group size, but the presence of countin is not necessary for CF50 to bind to cells, and CF50 is not necessary for countin to bind to cells. Our working hypothesis is that a signal transduction pathway activated by countin binding to cells modulates a signal transduction pathway activated by CF50 binding to cells and vice versa and that these two pathways can be distinguished by their effects on cAMP signal transduction.
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U2 - 10.1074/jbc.M309101200
DO - 10.1074/jbc.M309101200
M3 - Article
C2 - 14557265
AN - SCOPUS:0347362817
SN - 0021-9258
VL - 278
SP - 52262
EP - 52272
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 52
ER -