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The myeloid zinc finger gene, MZF-1, regulates the CD34 promoter in vitro
JF Morris, FJ Rauscher , B Davis, M Klemsz, D Xu, D Tenen and R Hromas
Medical College of Wisconsin, Milwaukee, USA.
MZF-1 is a C2H2 zinc finger gene encoding a putative transcriptional
regulator of myeloid differentiation. The MZF-1 protein contains 13 C2H2
zinc fingers arranged in bipartite DNA binding domains containing zinc
fingers through 4 and, in the carboxy-terminus, 5 through 13. We previously
identified the DNA consensus binding site recognized by the two DNA binding
domains. To assess the transcription regulatory function of MZF-1, the
full-length MZF-1 coding region was fused to the DNA binding domain of the
yeast transactivator GAL4. The expression vector was cotransfected with the
chloramphenicol acetyl transferase (CAT) reporter gene regulated by the
thymidine kinase promoter containing GAL4 DNA binding sites into NIH 3T3,
293, K562, and Jurkat cell lines. MZF-1 represses CAT reporter gene
expression via GAL4 binding sites in the nonhematopoietic cell lines NIH
3T3 and 293. In contrast, MZF-1 activates CAT reporter gene expression in
the hematopoietic cell lines K562 and Jurkat. The MZF-1 binding sites are
present in the promoters of several genes expressed during myeloid
differentiation, including the CD34 promoter. MZF-1 transcriptional
regulation of this physiologically relevant promoter was assessed in both
hematopoietic and nonhematopoietic cell lines. Recombinant MZF-1 protein
specifically binds to the consensus binding sites in the CD34 promoter in
mobility shift assays. MZF-1 expression vectors were cotransfected with the
luciferase reporter plasmids regulated by the CD34 promoter into both
nonhematopoietic and hematopoietic cell lines. As with the heterologous DNA
binding domain, MZF-1 represses reporter gene expression in
nonhematopoietic cell lines and activates expression in hematopoietic cell
lines. Activation of CD34 expression in hematopoietic cell lines is
dependent on the presence of intact MZF-1 binding sites. The cell
type-specific regulation of the CD34 promoter by MZF-1 suggests the
presence of tissue-specific regulators/adapters or differential MZF-1
modifications that determine MZF-1 transcriptional regulatory function.
Volume 86,
Issue 10,
pp. 3640-3647,
11/15/1995
Copyright © 1995 by The American Society of Hematology
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