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Blood, 1 December 2001, Vol. 98, No. 12, pp. 3283-3289
HEMATOPOIESIS
Notch1 inhibits differentiation of hematopoietic
cells by sustaining GATA-2 expression
Keiki Kumano,
Shigeru Chiba,
Kiyoshi Shimizu,
Tetsuya Yamagata,
Noriko Hosoya,
Toshiki Saito,
Tokiharu Takahashi,
Yoshio Hamada, and
Hisamaru Hirai
From the Department of Hematology and Oncology,
Graduate School of Medicine, University of Tokyo, Tokyo, Japan and the
National Institute of Basic Biology, Okazaki, Japan.
Notch signaling is involved in cell fate decisions in many systems
including hematopoiesis. It has been shown that expression of an
activated form of Notch1 (aNotch1) in 32D mouse myeloid progenitor
cells inhibits the granulocytic differentiation induced by granulocyte
colony-stimulating factor (G-CSF). Results of the current study show
that aNotch1, when expressed in F5-5 mouse erythroleukemia cells, also
inhibits erythroid differentiation. Comparison of the expression
levels of several transcription factors after stimulation for
myeloid and erythroid differentiation, in the presence or absence of
aNotch1, revealed that aNotch1 did not change its regulation pattern
with any of the transcription factors examined, except for GATA-2,
despite its inhibitory effect on differentiation. GATA-2 was
down-regulated when the parental 32D and F5-5 were induced to
differentiate into granulocytic and erythroid lineages, respectively.
In these induction procedures, however, the level of GATA-2 expression
was sustained when aNotch1 was expressed. To ascertain whether
maintenance of GATA-2 is required for the Notch-induced inhibition of
differentiation, the dominant-negative form of GATA-3 (DN-GATA), which
acted also against GATA-2, or transcription factor PU.1, which was
recently shown to be the repressor of GATA-2, was introduced
into aNotch1-expressing 32D (32D/aNotch1) cells that do not express
GATA family proteins other than GATA2. Both DN-GATA and PU.1 reversed
the phenotype of 32D/aNotch1 inducing its differentiation when G-CSF
was added. Furthermore, enforced expression of HES-1, which is involved
in Notch signaling, delayed differentiation of 32D, and again this
phenotype was neutralized by DN-GATA. These results indicate that
GATA-2 activity is necessary for the Notch signaling in hematopoietic cells.
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