A human erythropoietin receptor gene mutant causing familial erythrocytosis is associated with deregulation of the rates of Jak2 and Stat5 inactivation
Mo. Arcasoy et al., A human erythropoietin receptor gene mutant causing familial erythrocytosis is associated with deregulation of the rates of Jak2 and Stat5 inactivation, EXP HEMATOL, 27(1), 1999, pp. 63-74
The erythropoietin receptor (EpoR) has been previously shown to contain a c
ytoplasmic C-terminal negative regulatory domain, experimental deletion or
mutation of which leads to increased sensitivity of expressing cells to the
effects erythropoietin (Epo), We have studied a naturally occurring C-term
inal truncation mutant of the human EpoR by stably transfecting the growth
factor-dependent hematopoietic tissue culture cell line 32D with expression
plasmids containing either the wildtype or mutant human EpoR cDNA, thus re
ndering the cells dependent on Epo for viability and proliferation, In Epo
dose-response assays, cells expressing the mutant EpoR displayed hyperrespo
nsiueness to Epo compared with cells expressing comparable numbers of the w
ild-type EpoR cultured in the presence of fetal bovine serum. We investigat
ed whether enhanced Epo sensitivity of cells expressing the truncated EpoR
is associated with alteration in Epo receptor-mediated activation of Stat5,
which could have a role in Epo-induced proliferation, Although maximal Sta
t5 activation in response to a given concentration of Epo was comparable in
32D cells expressing the wild-type or truncated EpoRs, the time course of
Epo-induced Stat5 activation was very different. Gel-mobility shift studies
revealed the presence of Stat5 DNA-binding activity in nuclear and cytopla
smic extracts of cells expressing the truncated EpoR for a significantly lo
nger time than that observed in similar extracts of cells expressing the wi
ld-type EpoR consistent with decreased rate of inactivation of Stat5 in cel
ls expressing the mutant EpoR, Epo-dependent tyrosine phosphorylation of bo
th Stat5 and Jak2 was also substantially prolonged in cells expressing the
truncated EpoR, These results suggest a role for Stat5 in regulation of Epo
-mediated cell growth and implicate altered kinetics of Epo-induced Jak2 an
d Stat5 activation in the pathogenesis of familial erythrocytosis associate
d with this naturally occurring EpoR gene mutation. (C) 1999 International
Society for Experimental Hematology. Published by Elsevier Science Inc.