Myb and Ets proteins cooperate to transactivate an early myeloid gene.
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.
Shapiro
L H
LH
Department of Experimental Oncology, St. Jude Children's Research Hospital, Memphis, Tennessee 38101, USA.
D000483:Alleles; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D018826:CD13 Antigens / Q000235:genetics*; D002460:Cell Line; D005786:Gene Expression Regulation; D006801:Humans; D007962:Leukocytes / Q000166:cytology; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D011401:Promoter Regions, Genetic; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D050783:Proto-Oncogene Proteins c-ets; D020598:Proto-Oncogene Proteins c-myb; D014157:Transcription Factors; D015533:Transcriptional Activation
C494041:ETS1 protein, human; D015345:Oligonucleotide Probes; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D020598:Proto-Oncogene Proteins c-myb; D014157:Transcription Factors; D018826:CD13 Antigens