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Prepublished online as a Blood First Edition Paper on December 19, 2002; DOI 10.1182/blood-2002-04-1039.
PHAGOCYTES
From the Division of Hematology/Oncology, Cedars-Sinai
Medical Center, Los Angeles, CA; the Burns and Allen Research
Institute, University of California at Los Angeles School of Medicine;
the Departments of Molecular and Experimental Medicine and Immunology,
The Scripps Research Institute, La Jolla, CA; and the Center for Sleep
Respiratory Disorders, Fukoka, Japan.
In the bone marrow of C/EBP Transcription factors play a major role in the
development of specific lineages in the hematopoietic
system.1,2 Mature cells of the myeloid lineage, including
peripheral blood monocytes, tissue macrophages, and neutrophilic
and eosinophilic granulocytes, derive from a common myeloid
precursor. Of the numerous transcription factors involved in
myelopoiesis, C/EBP In the 2 murine models for PU.1 deficiency, mice either die at late
gestation or survive for approximately 48 hours after birth and succumb
to septicemia.5,6 Both models have defects in the myeloid
and lymphoid lineages. A lack of mature macrophages, neutrophils,
dendritic cells, osteoclasts, B cells, and T cells is
observed.5-7 Antibiotic treatment allows mice that survive birth to live up to 2 weeks, during which time some cells with the
characteristics of neutrophils develop by day 3.6 These neutrophils appear normal by morphology and express neutrophil markers
such as Gr-1 and chloroacetate esterase, but they fail to mature
completely as indicated by the lack of secondary granule gene
expression.8 In addition, neutrophils from PU.1 knockout mice are functionally impaired with defects in superoxide production, bacterial uptake, and killing.8 Restoration of
PU.1 gene expression in a myeloid cell line derived from the
embryonic livers of these mice reinstated expression of the secondary
granule genes, and functions of normal terminal neutrophil maturation
were acquired.9
Mice lacking the C/EBP Like C/EBP Phenotypic and functional defects of neutrophils and eosinophils
observed in C/EBP Together with in vitro studies of gene expression, these murine models
aid in the identification of target genes for these transcription
factors. A number of myeloid-specific genes contain functional C/EBP-
and PU.1 binding sites in their promoters, making them potential
targets for C/EBP C/EBP Transfections and generation of stable cell lines
Expression vectors and promoter-reporter gene constructs The cDNA encoding human C/EBP 30 was
subcloned from pcDNAI-C/EBP 30 into pcDNA3 (Invitrogen,
Carlsbad, CA).27 The pcDNA3 expression vector containing
the human C/EBP 32 isoform was a generous gift from Dr K. Xanthopoulos (Aurora, San Diego, CA). The pMSV-C/EBP (rat)
expression vector was kindly provided by Dr A. Friedman (Johns Hopkins
University, Baltimore, MD) and the expression vectors
pXM-GATA-128 and pMT2-FOG (friend of gata) (murine)29 were kind gifts from Dr S. Orkin (Harvard
University, Boston, MA). The human major basic protein gene
(hMBP) luciferase reporter construct containing sequence
between positions nucleotide (nt) 117 to +47 of the hMBP
P2 promoter region was described previously.30
Construction of the zinc-inducible C/EBP 32 vector pMT- 32 was described previously.31 To
construct the zinc-inducible rat C/EBP expression vector, a
1.1-kilobase pair (kbp) NcoI C/EBP cDNA fragment was
blunt-ended with Klenow and subcloned into EcoRV-cut pMT-CB6+ (kindly provided by F. Rauscher).32 The pCMVSPORT
vectors expressing human C/EBP 32,
C/EBP 30, and C/EBP were generated as described
previously.33
RNA isolation and analysis Total RNA was isolated by lysis of cells in TRIzol reagent as described by the manufacturer (Gibco/BRL). For reverse transcription-polymerase chain reaction (RT-PCR) analysis, total RNA was treated with RNase-free DNaseI (Promega, Madison, WI) and synthesized into cDNA with Moloney murine leukemia virus (MMLV) reverse transcriptase in a 50 µL volume as described by the manufacturer (Gibco/BRL). For transiently transfected cells, the entire RNA sample was reverse transcribed; for stably transformed cells, 2 µg RNA was used. PCR was performed with 1 µL cDNA per reaction using HotStar Taq polymerase (Qiagen, Chatsworth, CA). After a 15-minute denaturing step at 94°C, reactions were 94°C for 30 seconds, 55°C, 56°C, or 58°C for 30 seconds, and 72°C for 30 seconds. Cycle numbers for each primer set are described in the figure legends. Products were electrophoresed on 2% agarose gels and blotted for Southern analysis as described previously.17 Primers used for PCR and Southern blot hybridization were described previously15 or are summarized in Table 1. Each primer pair used for RT-PCR was designed to span 1 or more introns; therefore, amplification of contaminating genomic DNA would result in a product much larger than expected for amplification of the cDNA. A region 1 probe (N-terminal transcriptional activation domain, more than 90% similar to murine) of the human C/EBP gene was used to for Northern
hybridization.34
Quantitative real-time-PCR (QRT-PCR) for major basic protein
(MBP) expression was performed using a second primer set that spanned
an intron, as described in Table 1. Triplicate reactions for each cDNA
were set up as described in the previous paragraph, and SYBR
Green I (Molecular Probes, Eugene, OR) was added at a 1:60 000
dilution. After denaturing the template 15 minutes at 95°C, a 4-step
PCR reaction of 95°C for 30 seconds, 60°C for 30 seconds, 72°C
for 30 seconds, and 80°C for 20 seconds was performed. At the last
step, measurement of incorporated SYBR Green I was performed at 2°C
below the empirically determined melting temperature for the PCR
product. This melted all potential nonspecific products while
maintaining the specific product and ensuring that nonspecific products
were not detected. Gel electrophoresis confirmed that nonspecific
amplification was negligible. The balance of the cDNAs was determined
by quantifying the relative levels of 18S RNA in the samples. This was
performed with a TaqMan probe (FAM-agcaggcgcgcaaattaccc-TAMRA) and
amplification using TaqMan Universal PCR Master Mix (PE Biosystems, Foster City, CA). The nucleotide sequence of the PCR primers for 18S
was 5'-aaacggctaccacatccaag-3' (18S-F) and 5'-cctccaatggatcctcgtta-3' (18S-R). The threshold cycle (Ct) for each was determined,
and the expression levels of MBP were normalized to 18S. The fold change (FC) of the expression vector (v)-transfected samples compared with the empty vector (ev)-transfected control was determined by the
following equation: FC = 2 For Northern blot analysis, total RNA was electrophoresed through 1%
agarose/formaldehyde gels and was transferred to Hybond N+ membranes
(Amersham Life Sciences, Arlington Heights, IL). Probes were
synthesized with a Strip-EZ random priming kit (Ambion, Austin, TX)
with the incorporation of 5'-[ Protein isolation and analysis For the stable cell lines, total cell protein was prepared and Western blot analysis was performed as described previously.17 For transient transfections, total cell protein was prepared from the organic phase of the Trizol lysate as described by the manufacturer (Gibco/BRL). The antiserum against the amino-terminal half of C/EBP was described
previously.35 Commercially available antibodies against
C/EBP (C-22), C/EBP (14AA), C/EBP (C19), and C/EBP (M17)
were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). All
antibodies were used at a final concentration of 0.2 µg/mL. The
primary antibody was detected with a donkey-antirabbit horseradish
peroxidase (HRP) conjugate (Amersham Life Sciences) diluted 1:5000.
Antigen-antibody complexes were visualized using the Supersignal
chemiluminescence kit (Pierce, Rockford, IL) and exposure to Kodak
XO-Mat film.
Electrophoresis mobility shift assays COS-1 cells were transfected with empty or PU.1 expression vector as described in "Transfections and generation of stable cell lines." For total cell extract, cells were washed with phosphate-buffered saline (PBS) and were lysed in NP-40 lysis buffer (50 mM Tris-HCl, pH 7.4, 150 mM NaCl, 0.5% NP-40) containing Complete protease inhibitor cocktail (Roche Applied Science, Indianapolis, IN). Double-stranded oligonucleotides were end labeled with 32P- adenosine triphosphate (ATP; Dupont/NEN) using T4
polynucleotide kinase (Invitrogen) as described by the manufacturer.
Oligonucleotide sequences (sense-strand) were: (1) wild-type consensus
PU.1 5'-gggctgcttgaggaagtataagaat-3'; (2) mutant consensus
PU.1 5'-gggctgcttgagagagtataagaat-3'; (3) wild-type MBP
PU.1 5'-tctccctgggggaagttcctccaaggcc-3'; and
(4) mutant MBP PU.1
5'-tctccctgggcaaagtttgtccaaggcc-3'. Core
sequences are highlighted as underlined text. Electrophoresis mobility
shift assays (EMSAs) were performed as described
previously.34
Absence of secondary granule gene mRNA expression in the
C/EBP
The bone marrow of C/EBP
Generation of stable NIH 3T3 cell lines capable of zinc-inducible
expression of C/EBP or C/EBP in several leukemia
cell lines promotes granulocytic differentiation.31,37
This, in turn, leads to induction of a number of myeloid-specific genes that may or may not be direct targets of the transcription factors. In
addition, hematopoietic cell lines express transcription factors important in the regulation of myeloid-specific genes, thus making it
difficult to discern the contribution of these other transcription factors in cooperative activation. Myeloid-specific gene expression was
induced in avian fibroblasts by the coexpression of exogenous NFM
(nuclear factor myeloid, also known as avian C/EBP ) and
MYB.38,39 This occurs in the absence of differentiation
and eliminates the induction of genes that occurs during this process.
We reasoned that a similar phenomenon would occur in a mammalian
fibroblast cell line such as NIH 3T3. To determine whether secondary
granule genes are direct targets of C/EBP
Activation of neutrophil secondary granule gene expression in NIH 3T3 To determine the ability of C/EBP and C/EBP to activate gene
expression of secondary granule genes, the stable cell lines were
incubated in the absence or presence of ZnSO4 for 48 hours. Total RNA was subjected to RT-PCR analysis for genes that are deficient
in the bone marrow of the C/EBP -null mice. Both C/EBP and
C/EBP induced expression of MCLP, NGAL, and NC (Figure 3C). Interestingly, at 30 to 33 cycles, B9 was detected only in the C/EBP -expressing cell line (Figure 3C); however, at 35 cycles, low
levels of B9 were observed in the C/EBP stable line (data not
shown). The induction of B9 by C/EBP was detected by Northern blot
analysis as well (Figure 3B). These results demonstrated that using
both RT-PCR and Northern assays, we could detect the expression of
neutrophil-specific genes in a nonhematopoietic cell system.
Differential induction of neutrophil secondary granule genes mediated by C/EBP family members The C/EBP family members are all capable of binding the same DNA site and activating the same promoters in promoter-reporter assays. To compare the ability of the different C/EBP family members to activate secondary granule gene expression in NIH 3T3, cells were transiently transfected with expression vectors either for C/EBP 32,
C/EBP 30, C/EBP , C/EBP , or C/EBP . RT-PCR
analysis for the genes MCLP, NC, or B9 was performed (Figure 4, top
panels). Western blot analysis of the
proteins extracted from the organic phase of the TRIzol lysate prepared
from the transfected cells was performed to demonstrate that the
respective proteins were expressed (Figure 4, lower panels). A cDNA
prepared from 32Dcl3 cells served as a positive control (+) and a cDNA
from empty vector transfected cells was used as a negative control
( ).
Both C/EBP Cooperative transcriptional activation of MBP gene expression in
NIH 3T3 cells by C/EBP in NIH 3T3 cells alone was unable to
induce MBP expression efficiently (Figure
5). Previous studies demonstrated that
GATA-1 and C/EBP synergistically activate the human MBP-P2
promoter.41 Because MBP expression is deficient in the
bone marrow cells of the C/EBP![]() / mouse (Figure 2) but
not in the C/EBP![]() / mouse (data not shown), we
cotransfected C/EBP with GATA-1. A slight increase in MBP levels
over either C/EBP or GATA-1 alone was observed (Figure 5A). We
hypothesized that an additional factor was required for the induction
of high levels of MBP expression. Alignment of the nucleotide sequence
of the murine and human MBP promoter regions revealed
conservation of the C/EBP and GATA-1 sites (Figure
6). In addition, 2 GGAA core sequence
motifs (1 and 2) in a purine-rich region of the promoters indicated a
potential PU.1 binding site (Figure 6). The PU.1 core element 2 was
perfectly conserved between the murine and human promoters, but core
element 1 was not. This suggested the possibility that PU.1 may be
involved in the regulation of MBP gene expression.
To determine whether PU.1 cooperates with C/EBP
Mutation of the conserved PU.1 site in the human MBP-P2 promoter abolishes cooperative transcriptional activation with PU.1 To determine which core element in the conserved PU.1 site was required for the cooperative activation of the human MBP promoter, either core 1 (mutant [mt] 1), core 2 (mt 2), or both (mt 1-2) core sequences were mutated in the reporter construct pMBP(-117)-LUC. Wild-type or mutant reporter constructs were cotransfected with expression vectors for C/EBP , GATA-1, or
PU.1 alone or in various combinations. The mt 2 and mt 1-2 reporter
constructs lost response to PU.1, but not C/EBP or GATA-1, compared
with the wild-type or mt 1 reporter constructs (Figure
8). In addition, the synergistic activation by all 3 factors was abrogated in the mt 2 and mt 1-2 reporter constructs (Figure 8). These results indicate that the highly
conserved core 2 sequence is required for activation of the
MBP promoter by PU.1. In transfections with expression
vectors for C/EBP or GATA-1, the mt 2 and mt 1-2 reporter constructs activated less efficiently. This may be attributed to the loss of
binding by ETS-like factors found in NIH 3T3 cells to the mutated promoters whereas their binding would occur in the wild-type and mt 1 promoter constructs.
The loss of response to PU.1 was predicted to result from the loss of
PU.1 binding to the promoter. Electrophoretic mobility shift assay
(EMSA) revealed that PU.1 expressed in COS-1 cells was able to bind to
its consensus site and that this binding was blocked by the addition of
anti-PU.1 antibody (Figure 9, left panel). The binding of PU.1 to the
consensus site was competed by the unlabeled wild-type consensus and
the wild-type MBP PU.1 site, but not when the PU.1 sites were mutated
in either of these oligonucleotides (Figure 9, middle panel). Finally,
PU.1 binding to the wild-type MBP PU.1 site was detected in COS-1
extracts expressing PU.1, and this binding was blocked by the addition of anti-PU.1 antibody (Figure 9, right panel). These data demonstrate the binding of PU.1 to the predicted site in the MBP promoter and,
together with the reporter construct data, indicate that the loss of
promoter activation by PU.1 results from a lack of PU.1 binding to the
mutated core 2 site.
Eosinophil granule gene expression is absent in PU.1-deficient cells To determine the role of PU.1 in the expression of eosinophil granule genes, levels of MBP and EPX mRNA were determined in a myeloid cell line derived from the embryonic liver cells of PU.1-deficient mice.8,9 Like the bone marrow of C/EBP -deficient mice,
these cells lacked expression of neutrophil secondary granule mRNAs
such as LF, NG,8 and B9 (Figure 10, lanes
1-3), but they expressed primary
granule mRNAs including MPO and NE8 (Figure 10, lanes
1-3). Expression of the eosinophil granule mRNAs MBP and EPX was
deficient in the PU.1-null cell line (Figure 10, lane 3). When PU.1
expression was reinstated by retroviral transduction, neutrophil9 and eosinophil granule gene expression was
restored (Figure 10, lanes 4-5). This was not observed on the
restoration of M-CSF receptor expression by retroviral transduction
(Figure 10, lane 6). These results indicate that both C/EBP and PU.1
are important for the expression of eosinophil granule proteins in the mouse.
Studies on neutrophil secondary granule gene expression
demonstrate an important role for the C/EBP family in granule gene transcriptional regulation. The LF promoter possesses a
C/EBP binding site near its transcriptional start site that C/EBP
proteins bind to and activate transcription.15,42
The overlapping expression patterns of the C/EBP family members during
granulopoiesis obscure the roles the different C/EBP family members
play in the in vivo regulation of these genes. Neutrophil secondary
granule gene expression is severely impaired in
C/EBP Granulocytes of C/EBP Stable and transient expression of the 2 C/EBP family members most
critical for granulocytic differentiation, C/EBP If both C/EBP Both PU.1 and GATA-1 interact at the protein level and reciprocally inhibit the transcription- and differentiation-inducing functions of each other.54-58 These studies suggest this interaction is important for the decision of stem cells to differentiate down the erythroid versus the myeloid lineage. Our study demonstrates that the consequences of PU.1 and GATA-1, interacting with their adjacent binding sites on a promoter of a target gene, result in the activation rather than the inhibition of gene expression. Similarly, PU.1 and GATA-1 cooperatively stimulated activity of the mast cell-specific intronic enhancer of IL-4.59 In summary, we demonstrated the ability of C/EBP family members to
differentially activate granulocytic secondary granule genes in a
nonhematopoietic cell line. This suggests that, in addition to
C/EBP
We thank Kleanthis Xanthopolous and Julie Lekstrom-Himes for
sharing the C/EBP
Submitted April 4, 2002; accepted November 27, 2002.
Prepublished online as Blood First Edition Paper, December 19, 2002; DOI 10.1182/blood-2002-04-1039.
Supported by National Institutes of Health grants CA26038-20 (H.P.K.) and DK54938 (B.E.T.), the Joseph Troy Leukemia Fund, the Horn Foundation, the Lymphoma Research Foundation of America, and the C. and H. Koeffler Fund. A.F.G. is a recipient of a Lymphoma Research Foundation of America Fellowship. H.P.K. holds the Mark Goodson endowed chair for Cancer Research and is a member of the Jonsson Cancer Center.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: Adrian F. Gombart, Division of Hematology/Oncology, Cedars-Sinai Medical Center, Davis Bldg 5019, Los Angeles, CA 90048; e-mail: gombarta{at}csmc.edu.
1.
Shivdasani RA, Orkin SH.
The transcriptional control of hematopoiesis [see comments].
Blood.
1996;87:4025-4039
2.
Tenen DG, Hromas R, Licht JD, Zhang DE.
Transcription factors, normal myeloid development, and leukemia.
Blood.
1997;90:489-519
3.
Yamanaka R, Barlow C, Lekstrom-Himes J, et al.
Impaired granulopoiesis, myelodysplasia, and early lethality in CCAAT/enhancer binding protein epsilon-deficient mice.
Proc Natl Acad Sci U S A.
1997;94:13187-13192 4. Tanaka T, Akira S, Yoshida K, et al. Targeted disruption of the NF-IL6 gene discloses its essential role in bacteria killing and tumor cytotoxicity by macrophages. Cell. 1995;80:353-361[CrossRef][Medline] [Order article via Infotrieve].
5.
Scott EW, Simon MC, Anastasi J, Singh H.
Requirement of transcription factor PU.1 in the development of multiple hematopoietic lineages.
Science.
1994;265:1573-1577 6. McKercher SR, Torbett BE, Anderson KL, et al. Targeted disruption of the PU.1 gene results in multiple hematopoietic abnormalities. EMBO J. 1996;15:5647-5658[Medline] [Order article via Infotrieve]. 7. Tondravi MM, McKercher SR, Anderson K, et al. Osteopetrosis in mice lacking haematopoietic transcription factor PU.1. Nature. 1997;386:81-84[CrossRef][Medline] [Order article via Infotrieve].
8.
Anderson KL, Smith KA, Pio F, Torbett BE, Maki RA.
Neutrophils deficient in PU.1 do not terminally differentiate or become functionally competent.
Blood.
1998;92:1576-1585
9.
Anderson KL, Smith KA, Perkin H, et al.
PU.1 and the granulocyte and macrophage colony-stimulating factor receptors play distinct roles in late-stage myeloid cell differentiation.
Blood.
1999;94:2310-2318
10.
Wang ND, Finegold MJ, Bradley A, et al.
Impaired energy homeostasis in C/EBP alpha knockout mice.
Science.
1995;269:1108-1112
11.
Flodby P, Barlow C, Kylefjord H, Ahrlund-Richter L, Xanthopoulos KG.
Increased hepatic cell proliferation and lung abnormalities in mice deficient in CCAAT/enhancer binding protein alpha.
J Biol Chem.
1996;271:24753-24760
12.
Zhang DE, Zhang P, Wang ND, et al.
Absence of granulocyte colony-stimulating factor signaling and neutrophil development in CCAAT enhancer binding protein alpha-deficient mice.
Proc Natl Acad Sci U S A.
1997;94:569-574
13.
Zhang P, Iwama A, Datta MW, et al.
Upregulation of interleukin 6 and granulocyte colony-stimulating factor receptors by transcription factor CCAAT enhancer binding protein
14.
Lekstrom-Himes J, Xanthopoulos KG.
CCAAT/enhancer binding protein epsilon is critical for effective neutrophil-mediated response to inflammatory challenge.
Blood.
1999;93:3096-3105
15.
Verbeek W, Lekstrom-Himes J, Park DJ, et al.
Myeloid transcription factor C/EBP
16.
Lekstrom-Himes JA, Dorman SE, Kopar P, Holland SM, Gallin JI.
Neutrophil-specific granule deficiency results from a novel mutation with loss of function of the transcription factor CCAAT/enhancer binding protein epsilon.
J Exp Med.
1999;189:1847-1852
17.
Gombart AF, Shiohara M, Kwok SH, et al.
Neutrophil-specific granule deficiency: homozygous recessive inheritance of a frameshift mutation in the gene encoding transcription factor CCAAT/enhancer binding protein-epsilon.
Blood.
2001;97:2561-2567 18. Gombart AF, Koeffler HP. Neutrophil specific granule deficiency and mutations in the gene encoding transcription factor C/EBP(epsilon). Curr Opin Hematol. 2002;9:36-42[CrossRef][Medline] [Order article via Infotrieve].
19.
Johnston JJ, Rintels P, Chung J, et al.
Lactoferrin gene promoter: structural integrity and nonexpression in HL60 cells.
Blood.
1992;79:2998-3006
20.
Nuchprayoon I, Meyers S, Scott LM, et al.
PEBP2/CBF, the murine homolog of the human myeloid AML1 and PEBP2 beta/CBF beta proto-oncoproteins, regulates the murine myeloperoxidase and neutrophil elastase genes in immature myeloid cells.
Mol Cell Biol.
1994;14:5558-5568 21. Hohaus S, Petrovick MS, Voso MT, et al. PU.1 (Spi-1) and C/EBP alpha regulate expression of the granulocyte macrophage colony-stimulating factor receptor alpha gene. Mol Cell Biol. 1995;15:5830-5845[Abstract].
22.
Smith LT, Hohaus S, Gonzalez DA, Dziennis SE, Tenen DG.
PU.1 (Spi-1) and C/EBP alpha regulate the granulocyte colony- stimulating factor receptor promoter in myeloid cells.
Blood.
1996;88:1234-1247
23.
Sturrock A, Franklin KF, Hoidal JR.
Human proteinase-3 expression is regulated by PU.1 in conjunction with a cytidine-rich element.
J Biol Chem.
1996;271:32392-32402 24. Zhang DE, Hohaus S, Voso MT, et al. Function of PU.1 (Spi-1), C/EBP, and AML1 in early myelopoiesis: regulation of multiple myeloid CSF receptor promoters. Curr Top Microbiol Immunol. 1996;211:137-147[Medline] [Order article via Infotrieve].
25.
Iwama A, Zhang P, Darlington GJ, et al.
Use of RDA analysis of knockout mice to identify myeloid genes regulated in vivo by PU.1 and C/EBP 26. Schwarze PE, Johnsen NM, Samuelsen JT. et al. The use of isolated lung cells in in vitro pulmonary toxicology: studies of DNA damage, apoptosis and alteration of gene expression. Cent Eur J Public Health. 1996;4(suppl):6-10[Medline] [Order article via Infotrieve]. 27. Park DJ, Chumakov AM, Miller CW, Pham EY, Koeffler HP. p53 transactivation through various p53-responsive elements. Mol Carcinog. 1996;16:101-108[CrossRef][Medline] [Order article via Infotrieve]. 28. Tsai SF, Martin DI, Zon LI, et al. Cloning of cDNA for the major DNA-binding protein of the erythroid lineage through expression in mammalian cells. Nature. 1989;339:446-451[CrossRef][Medline] [Order article via Infotrieve]. 29. Tsang AP, Visvader JE, Turner CA, et al. FOG, a multitype zinc finger protein, acts as a cofactor for transcription factor GATA-1 in erythroid and megakaryocytic differentiation. Cell. 1997;90:109-119[CrossRef][Medline] [Order article via Infotrieve].
30.
Yamaguchi Y, Ackerman SJ, Minegishi N, et al.
Mechanisms of transcription in eosinophils: GATA-1, but not GATA-2, transactivates the promoter of the eosinophil granule major basic protein gene.
Blood.
1998;91:3447-3458 31. Park DJ, Chumakov AM, Vuong PT, et al. CCAAT/enhancer binding protein epsilon is a potential retinoid target gene in acute promyelocytic leukemia treatment [see comments]. J Clin Invest. 1999;103:1399-1408[Medline] [Order article via Infotrieve]. 32. Cook DM, Hinkes MT, Bernfield M, Rauscher FJ. Transcriptional activation of the syndecan-1 promoter by the Wilms' tumor protein WT1. Oncogene. 1996;13:1789-1799[Medline] [Order article via Infotrieve].
33.
Tang JG, Koeffler HP.
Structural and functional studies of CCAAT/enhancer-binding protein epsilon.
J Biol Chem.
2001;276:17739-17746
34.
Gombart AF, Hofmann WK, Kawano S, et al.
Mutations in the gene encoding the transcription factor CCAAT/enhancer binding protein alpha in myelodysplastic syndromes and acute myeloid leukemias.
Blood.
2002;99:1332-1340 35. Chumakov AM, Grillier I, Chumakova E, et al. Cloning of the novel human myeloid cell-specific C/EBP-epsilon transcription factor. Mol Cell Biol. 1997;17:1375-1386[Abstract].
36.
Rosenberg HF, Gallin JI.
Neutrophil-specific granule deficiency includes eosinophils.
Blood.
1993;82:268-273
37.
Radomska HS, Huettner CS, Zhang P, et al.
CCAAT/enhancer binding protein alpha is a regulatory switch sufficient for induction of granulocytic development from bipotential myeloid progenitors.
Mol Cell Biol.
1998;18:4301-4314
38.
Ness SA, Kowenz-Leutz E, Casini T, Graf T, Leutz A.
Myb and NF-M: combinatorial activators of myeloid genes in heterologous cell types.
Genes Dev.
1993;7:749-759 39. Burk O, Mink S, Ringwald M, Klempnauer KH. Synergistic activation of the chicken mim-1 gene by v-myb and C/EBP transcription factors. EMBO J. 1993;12:2027-2038[Medline] [Order article via Infotrieve].
40.
Verbeek W, Gombart AF, Chumakov AM, et al.
C/EBP
41.
Yamaguchi Y, Nishio H, Kishi K, Ackerman SJ, Suda T.
C/EBP
42.
Khanna-Gupta A, Zibello T, Simkevich C, Rosmarin AG, Berliner N.
Sp1 and C/EBP are necessary to activate the lactoferrin gene promoter during myeloid differentiation.
Blood.
2000;95:3734-3741
43.
Hu HM, Baer M, Williams SC, Johnson PF, Schwartz RC.
Redundancy of C/EBP alpha, -beta, and -delta in supporting the lipopolysaccharide-induced transcription of IL-6 and monocyte chemoattractant protein-1.
J Immunol.
1998;160:2334-2342
44.
Williams SC, Du Y, Schwartz RC, et al.
C/EBP
45.
Wang W, Wang X, Ward AC, Touw IP, Friedman AD.
C/EBP
46.
Freytag SO, Paielli DL, Gilbert JD.
Ectopic expression of the CCAAT/enhancer-binding protein alpha promotes the adipogenic program in a variety of mouse fibroblastic cells.
Genes Dev.
1994;8:1654-1663
47.
Wu Z, Xie Y, Bucher NL, Farmer SR.
Conditional ectopic expression of C/EBP beta in NIH-3T3 cells induces PPAR gamma and stimulates adipogenesis.
Genes Dev.
1995;9:2350-2363
48.
Hollenberg SM, Cheng PF, Weintraub H.
Use of a conditional MyoD transcription factor in studies of MyoD trans-activation and muscle determination.
Proc Natl Acad Sci U S A.
1993;90:8028-8032 49. Teitell MA, Thompson AD, Sorensen PH, et al. EWS/ETS fusion genes induce epithelial and neuroectodermal differentiation in NIH 3T3 fibroblasts. Lab Invest. 1999;79:1535-1543. 50. Khanna-Gupta A, Zibello T, Kolla S, Neufeld EJ, Berliner N. CCAAT displacement protein (CDP/cut) recognizes a silencer element within the lactoferrin gene promoter. Blood. 1997;90:2784-2795.
51.
Scott LM, Civin CI, Rorth P, Friedman AD.
A novel temporal expression pattern of three C/EBP family members in differentiating myelomonocytic cells.
Blood.
1992;80:1725-1735
52.
Du J, Savage MP, Dekoter R, et al.
Impaired eosinophilopoiesis in PU.1 deficient mice [abstract].
Blood.
1998;92:191
53.
van Dijk TB, Caldenhoven E, Raaijmakers JA, et al.
The role of transcription factor PU.1 in the activity of the intronic enhancer of the eosinophil-derived neurotoxin (RNS2) gene.
Blood.
1998;91:2126-2132
54.
Nerlov C, Querfurth E, Kulessa H, Graf T.
GATA-1 interacts with the myeloid PU.1 transcription factor and represses PU.1-dependent transcription.
Blood.
2000;95:2543-2551
55.
Zhang P, Zhang X, Iwama A, et al.
PU.1 inhibits GATA-1 function and erythroid differentiation by blocking GATA-1 DNA binding.
Blood.
2000;96:2641-2648
56.
Zhang P, Behre G, Pan J, et al.
Negative cross-talk between hematopoietic regulators: GATA proteins repress PU.1.
Proc Natl Acad Sci U S A.
1999;96:8705-8710
57.
Rekhtman N, Radparvar F, Evans T, Skoultchi AI.
Direct interaction of hematopoietic transcription factors PU.1 and GATA-1: functional antagonism in erythroid cells.
Genes Dev.
1999;13:1398-1411 58. Yamada T, Kihara-Negishi F, Yamamoto H, et al. Reduction of DNA binding activity of the GATA-1 transcription factor in the apoptotic process induced by overexpression of PU.1 in murine erythroleukemia cells. Exp Cell Res. 1998;245:186-194[CrossRef][Medline] [Order article via Infotrieve].
59.
Henkel G, Brown MA.
PU.1 and GATA: components of a mast cell-specific interleukin 4 intronic enhancer.
Proc Natl Acad Sci U S A.
1994;91:7737-7741
© 2003 by The American Society of Hematology.
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