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Volume 272, Number 8, Issue of February 21, 1997 pp. 4964-4969
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

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Activation of a Calcium-permeable Cation Channel CD20 Expressed in Balb/c 3T3 Cells by Insulin-like Growth Factor-I

(Received for publication, August 8, 1996, and in revised form, October 4, 1996)

From the Department of Cell Biology, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi 371, Japan

CD20 functions as a calcium-permeable cation channel. When expressed in Balb/c 3T3 cells, CD20 accelerates the G₁ progression induced by insulin-like growth factor-I (IGF-I). To further characterize how CD20 modulates the action of IGF-I, we investigated whether the activity of CD20 channel was affected by IGF-I. In quiescent cells expressing CD20, IGF-I increased cytoplasmic free calcium concentration, [Ca²⁺]_c, which was reversed by the removal of extracellular calcium. In contrast, IGF-I did not increase [Ca²⁺]_c in cells that did not express CD20. In perforated patch clamp recordings, addition of IGF-I to the bath solution augmented the Ca²⁺ permeability, which was reversed by anti-CD20 antibody. In cell-attached patch, calcium-permeable channel activity with unitary conductance of 7 picosiemens was detected, which was abolished by anti-CD20 antibody. The single channel activities were markedly enhanced when IGF-I was included in the pipette solution, whereas IGF-I added to the bath solution was ineffective. When cells were first exposed to pertussis toxin, activation of the channel by IGF-I was blocked. Transfection of cDNA for Gip2, a constitutive active form of _i2, activated the CD20 channel. These results indicate that the CD20 channel is regulated by the IGF-I receptor by a mechanism involving pertussis toxin-sensitive G protein.

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