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. 1998 Mar 3;95(5):2061-6.
doi: 10.1073/pnas.95.5.2061.

CREB-binding protein cooperates with transcription factor GATA-1 and is required for erythroid differentiation

G A Blobel  1 , T Nakajima, R Eckner, M Montminy, S H Orkin
Affiliations

CREB-binding protein cooperates with transcription factor GATA-1 and is required for erythroid differentiation

G A Blobel et al. Proc Natl Acad Sci U S A. .

Abstract

The transcription factor GATA-1 coordinates multiple events during terminal erythroid cell maturation. GATA-1 participates in the transcription of virtually all erythroid-specific genes, blocks apoptosis of precursor cells, and controls the balance between proliferation and cell cycle arrest. Prior studies suggest that the function of GATA-1 is mediated in part through association with transcriptional cofactors. CREB-binding protein (CBP) and its close relative p300 serve as coactivators for a variety of transcription factors involved in growth control and differentiation. We report here that CBP markedly stimulates GATA-1's transcriptional activity in transient transfection experiments in nonhematopoietic cells. GATA-1 and CBP also coimmunoprecipitate from nuclear extracts of erythroid cells. Interaction mapping pinpoints contact sites to the zinc finger region of GATA-1 and to the E1A-binding region of CBP. Expression of a conditional form of adenovirus E1A in murine erythroleukemia cells blocks differentiation and expression of endogenous GATA-1 target genes, whereas mutant forms of E1A unable to bind CBP/p300 have no effect. Our findings add GATA-1, and very likely other members of the GATA family, to the growing list of molecules implicated in the complex regulatory network surrounding CBP/p300.

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Figures

Figure 1
Figure 1
(A) CBP stimulates activity of GATA-1(Δ63) in a dose-dependent manner using a synthetic reporter (M1α-GH). Numbers indicate amounts of transfected plasmid in μg. (B) CBP potentiates GATA-1(Δ63)-mediated activation of synthetic (M1α-GH) and natural (EKLF-GH) GATA-1 target promoters by with comparable efficiency. (C) CBP potentiates wtGATA-1 (Upper). wt E1A, but not E1AΔ2–36, inhibit CBP-mediated activation (Upper) without affecting expression of GATA-1 and CBP as determined by Western blot analysis (Lower).
Figure 2
Figure 2
Coimmunoprecipitation of GATA-1 and CBP. CBP was precipitated with rabbit anti-CBP serum or, as control, rabbit nonimmune serum (n.i.) containing equal amounts of total IgG. GATA-1 was precipitated with a rat mAb or an isotype-matched irrelevant control rat mAb. The secondary anti-rat IgG antibody used for Western blot analysis recognized the rat IgG2a used for the GATA-1 immunoprecipitation. i.p.: indicates antibodies used for immunoprecipitation.
Figure 3
Figure 3
GST pulldown experiments demonstrating interaction between the finger domain of GATA-1 and the E1A-binding region (E1A-BD). (A) Mapping of the domain of CBP that interacts with GATA-1. Numbers indicate amino acids present in the in vitro translated CBP. (B) Mapping of the region of GATA-1 that interacts with CBP. Disr.Nf bears a deletion spanning amino acids 200–248. GST-GATA-1Δf lacks both fingers, GST-f(GATA-1) represents the finger region alone. GATA-1 binding to itself served as positive control. Twenty percent of in vitro translated product was analyzed directly (input).
Figure 4
Figure 4
Activated E1A-ER blocks MEL differentiation. (A) Control MEL cells untreated (U) or treated with estradiol (E), DMSO (D), or both (D/E). Cells were stained with benzidine and May-Grunwald. (B) A MEL line expressing E1A-ER. Note the marked reduction of benzidine positive cells in the sample treated with DMSO and estradiol.
Figure 5
Figure 5
Activated E1A-ER causes a block in α- and β-globin gene induction. Northern analysis of control MEL cells, two E1A-ER expressing lines, and two lines constitutively expressing high levels of E1AΔ2–36 and E1AΔ38–67 treated with DMSO or estradiol. Blots were probed with a labeled β-globin fragment, stripped, and successively reprobed with α-globin and actin cDNA probes.
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References

    1. Orkin S H. Curr Opin Cell Biol. 1995;7:870–877. - PubMed
    1. Weiss M J, Orkin S H. Exp Hematol. 1995;23:99–107. - PubMed
    1. Fujiwara Y, Browne C P, Cunniff K, Goff S C, Orkin S H. Proc Natl Acad Sci USA. 1996;93:12355–12358. - PMC - PubMed
    1. Weiss M J, Orkin S H. Proc Natl Acad Sci USA. 1995;92:9623–9627. - PMC - PubMed
    1. Weiss M J, Keller G, Orkin S H. Genes Dev. 1994;8:1184–1197. - PubMed

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