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Cloning and characterization of fetal liver phosphatase 1, a nuclear
protein tyrosine phosphatase isolated from hematopoietic stem cells
M Dosil, N Leibman and IR Lemischka
Department of Molecular Biology, Princeton University, NJ 08544-1014, USA.
We report the isolation of cDNAs encoding protein tyrosine phosphatases
(PTPs) from highly purified hematopoietic stem cell populations. One such
cDNA encodes a novel PTP, designated fetal liver phosphatase 1 (FLP1),
which consists of one PTP domain followed by a carboxy terminal domain of
160 amino acids. Northern blot and in situ hybridization analysis showed
that expression of FLP1 mRNA is restricted to thymus in 15.5-day-old and
17.5-day-old mouse embryos and to kidney and hematopoietic tissues in adult
mice. Furthermore, polymerase chain reaction-based analysis shows that FLP1
is expressed in hematopoietic stem cells as well as in more mature
hematopoietic cells. Peptide antisera against FLP1 immunoprecipitated a
48-kD protein that is localized in the nuclei of Ba/F3 lymphoid cells. We
have analyzed the effects of overexpressing either wild-type FLP1 or a
functionally inactive mutant of FLP1 in hematopoietic cells. In the
progenitor K562 cell line, cells ectopically expressing functional FLP1
differentiated normally to megakaryocytes after induction with
tetradecanoyl phorbol acetate (TPA). In contrast, when K562 transfectants
expressing an inactive mutant FLP1 protein were treated with TPA, the
characteristic cell spreading and substrate adhesion that accompany
megakaryocytic differentiation did not occur. We show that, in these cells,
the induction of the differentiation marker alphaIIb beta3 is not affected.
However, both constitutive and TPA-induced expression of alpha2 integrin, a
late megakaryocytic marker, are inhibited. These results suggest that the
expression of an inactive form of FLP1 affects late signaling events of
K562 megakaryocytic differentiation.
Volume 88,
Issue 12,
pp. 4510-4525,
12/15/1996
Copyright © 1996 by The American Society of Hematology
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