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The earliest progenitor cell committed to the
granulocyte/monocyte developmental pathway can be identified by the
appearance of a 150-kDa glycoprotein on the cell surface
(CD13/aminopeptidase N (CD13/APN), EC 3.4.11.2). A 455-base pair
genomic fragment from the CD13/APN gene containing a Myb
consensus-binding site as well as three potential Ets-binding sites was
found to regulate tissue-appropriate expression of reporter genes in
hematopoietic cell lines. Transactivation experiments with plasmids
expressing either a full-length or truncated Myb protein and the
full-length Ets-1 or Ets-2 protein demonstrated that these proteins
cooperate to positively regulate CD13/APN gene expression.
This cooperation is synergistic, as levels of transcriptional activity
produced by Myb and Ets in combination were higher than those expected
from a purely additive effect. Mutation of the Myb consensus-binding
site completely abolished CD13/APN promoter activity in
myeloid cells. Introduction of a dominant interfering Myb allele
disrupted the ability of endogenous c-Myb in myeloid cells to
transactivate the CD13/APN construct. Other myeloid cell-expressed Ets
family members (PU.1, Fli-1, and Elf-1) failed to produce a cooperative
transactivating effect when combined with the Myb expression construct.
These data contrast with previous studies indicating that full-length
c-Myb is unable to positively cooperate with Ets proteins in the
regulation of myeloid genes. Because intact c-Myb and Ets-2 proteins,
both endogenously expressed in myeloid cells, act synergistically to
transactivate the CD13/APN promoter, this gene may represent a
physiological target for dissection of the roles of these transcription
factors in normal and malignant myelopoiesis.
Volume 270,
Number 15,
Issue of April 14, pp. 8763-8771, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
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