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Blood, 1 July 2002, Vol. 100, No. 1, pp. 80-88
HEMATOPOIESIS
Transcriptional regulation of myeloid differentiation primary
response (MyD) genes during myeloid differentiation is mediated by
nuclear factor Y
Robert M. Tjin Tham Sjin,
Kandasamy Krishnaraju,
Barbara Hoffman, and
Dan A. Liebermann
From the Fels Institute for Cancer Research and
Molecular Biology and Department of Biochemistry, Temple University
School of Medicine, Philadelphia, PA.
To understand the molecular mechanism by which interleukin-6 (IL-6)
regulates myeloid differentiation primary response (MyD) genes at the
onset of M1 myeloid differentiation, we used JunB as a representative
MyD gene to isolate and characterize IL-6 responsive elements. An IL-6
responsive element was localized between  65 and 52 of the JunB
promoter (65/52 IL-6RE). By using antibody and oligonucleotide
competition assays in electrophoretic mobility shift assay experiments,
we have shown that the heterotrimeric transcription nuclear factor Y
(NF-Y) complex binds to this element. A dominant-negative form of
NF-YA, ectopically expressed in M1 cells, blocked NF-Y binding to the
65/52 IL-6RE and reduced induction of JunB by IL-6. Furthermore,
inhibition of NF-Y binding also reduced MyD gene induction by IL-6 and
dampened the IL-6-induced M1 differentiation program. These findings
are consistent with the observation that most MyD genes contain intact
NF-Y binding motifs in their promoter regions. In contrast to M1 cells,
during myeloid differentiation of bone marrow (BM), there was induction of NF-Y binding to the 65/52 IL-6RE. This induced binding can be
attributed to the observed induction of NF-YA protein expression and
may reflect the molecular mechanism that couples proliferation to
terminal differentiation of normal myeloblasts. Similar to M1 cells,
blocking NF-Y binding in BM resulted in a reduction in mature
macrophages. It can be concluded that NF-Y plays a role in the
transcriptional regulation of MyD genes and is required for optimum
myeloid differentiation.
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