Ligand- and kinase activity-independent cell survival mediated by the epidermal growth factor receptor expressed in 32D cells.
To investigate the intrinsic activities of the epidermal growth factor receptor and the role of its kinase domain in these functions within a cellular environment lacking endogenous ErbB protein expression, wild-type EGF receptor (WT-EGFR) and two kinase-impaired mutants, D813A and K721R, were expressed in 32D murine hematopoietic cells, a line which is normally dependent on interleukin 3 (IL3) for growth and survival. Addition of EGF in the absence of IL3 stimulates receptor autophosphorylation and, in the presence of serum, mitosis in cells expressing WT-EGFR, but not in cells expressing D813A or K721R. Unexpectedly, cells expressing WT-EGFR or K721R exhibited IL3-independent survival in the presence of fetal bovine serum; parental 32D cells and cells expressing D813A did not survive, apparently undergoing apoptosis in the absence of IL3, whether or not serum was present. Addition of EGF did not prevent the apoptosis of WT-EGFR or K721R cells in serum-free medium. Activation of Akt was not necessary to mediate the prosurvival activity of EGF receptor expression. These results suggest that the EGF receptor can mediate the prevention of apoptosis independently of both receptor-ligand binding and receptor kinase activity, and this activity is disrupted by the D813A mutation.
Ewald
Jonathan A
JA
Department of Biological Sciences, Vanderbilt University, Nashville, TN 37235, USA.
Wilkinson
John C
JC
Guyer
Cheryl A
CA
Staros
James V
JV
D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066246:ErbB Receptors / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000737:chemistry / Q000166:cytology*; D007377:Interleukin-3 / Q000494:pharmacology; D008024:Ligands; D051379:Mice; D008938:Mitosis / Q000187:drug effects; D009154:Mutation; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D017434:Protein Structure, Tertiary; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt
D007377:Interleukin-3; D008024:Ligands; D011518:Proto-Oncogene Proteins; D004815:Epidermal Growth Factor; D010770:Phosphotransferases; D066246:ErbB Receptors; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt