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Mol Cell Biol, July 1998, p. 3699-3707, Vol. 18, No. 7
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

AP1 Regulation of Proliferation and Initiation of Apoptosis in Erythropoietin-Dependent Erythroid Cells

Sarah M. Jacobs-Helber,1 Amittha Wickrema,2 Michael J. Birrer,3 and Stephen T. Sawyer1 *

Department of Pharmacology/Toxicology, Medical College of Virginia Campus of Virginia Commonwealth University, Richmond, Virginia 232981; Hematology/Oncology Section, University of Illinois at Chicago, Chicago, Illinois 606072; and Department of Cell and Cancer Biology, Medicine Branch, National Cancer Institute, National Institutes of Health, Rockville, Maryland 208503

Received 3 December 1997/Returned for modification 16 January 1998/Accepted 29 March 1998

The transcription factor AP1 has been implicated in the induction of apoptosis in cells in response to stress factors and growth factor withdrawal. We report here that AP1 is necessary for the induction of apoptosis following hormone withdrawal in the erythropoietin (EPO)-dependent erythroid cell line HCD57. AP1 DNA binding activity increased upon withdrawal of HCD57 cells from EPO. A dominant negative AP1 mutant rendered these cells resistant to apoptosis induced by EPO withdrawal and blocked the downregulation of Bcl-XL. JunB is a major binding protein in the AP1 complex observed upon EPO withdrawal; JunB but not c-Jun was present in the AP1 complex 3 h after EPO withdrawal in HCD57 cells, with a concurrent increase in junB message and protein. Furthermore, analysis of AP1 DNA binding activity in an apoptosis-resistant subclone of HCD57 revealed a lack of induction in AP1 DNA binding activity and no change in junB mRNA levels upon EPO withdrawal. In addition, we determined that c-Jun and AP1 activities correlated with EPO-induced proliferation and/or protection from apoptosis. AP1 DNA binding activity increased over the first 3 h following EPO stimulation of HCD57 cells, and suppression of AP1 activity partially inhibited EPO-induced proliferation. c-Jun but not JunB was present in the AP1 complex 3 h after EPO addition. These results implicate AP1 in the regulation of proliferation and survival of erythroid cells and suggest that different AP1 factors may play distinct roles in both triggering apoptosis (JunB) and protecting erythroid cells from apoptosis (c-Jun).

* Corresponding author. Mailing address: Department of Pharmacology/Toxicology, P.O. Box 980613, Richmond, VA 23298. Phone: (804) 828-7918. Fax: (804) 828-2117. E-mail: SSAWYER{at}GEMS.VCU.EDU.

Mol Cell Biol, July 1998, p. 3699-3707, Vol. 18, No. 7
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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