Dynamic GATA Factor Interplay at a Multicomponent Regulatory Region of the GATA-2 Locus

Martowicz, Melissa L.; Grass, Jeffrey A.; Boyer, Meghan E.; Guend, Hamza; Bresnick, Emery H.

doi:10.1074/jbc.m406038200

Cited by 81 publications

(111 citation statements)

References 51 publications

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“…1A). GATA-2 occupancy of the −3.9 site, first described in GATA-1-null G1E cells (12), occurs in multiple cell lines and primary cells, including lineage negative (Lin − ) hematopoietic progenitors from bone marrow (Fig. 1B).…”

Section: Sequence Conservation and Transcription Factor Occupancy Do Notmentioning

confidence: 82%

“…The −3.9 site harbors two inverted GATA motifs and contains canonical attributes of cis-regulatory elements, including DNase I hypersensitivity and GATA site-dependent enhancer activity in a transfection assay (9,12). Because these attributes are shared with one or more of the −2.8, −1.8, and +9.5 sites, which differ greatly in their importance in vivo, the contribution of the −3.9 site can only be ascertained by disruption of this site at the endogenous locus.…”

Section: Significancementioning

confidence: 99%

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Mechanism governing a stem cell-generating cis -regulatory element

Sanalkumar

Johnson

Gào

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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The unremitting demand to replenish differentiated cells in tissues requires efficient mechanisms to generate and regulate stem and progenitor cells. Although master regulatory transcription factors, including GATA binding protein-2 (GATA-2), have crucial roles in these mechanisms, how such factors are controlled in developmentally dynamic systems is poorly understood. Previously, we described five dispersed Gata2 locus sequences, termed the −77, −3.9, −2.8, −1.8, and +9.5 GATA switch sites, which contain evolutionarily conserved GATA motifs occupied by GATA-2 and GATA-1 in hematopoietic precursors and erythroid cells, respectively. Despite common attributes of transcriptional enhancers, targeted deletions of the −2.8, −1.8, and +9.5 sites revealed distinct and unpredictable contributions to Gata2 expression and hematopoiesis. Herein, we describe the targeted deletion of the −3.9 site and mechanistically compare the −3.9 site with other GATA switch sites. The −3.9 −/− mice were viable and exhibited normal Gata2 expression and steady-state hematopoiesis in the embryo and adult. We established a Gata2 repression/reactivation assay, which revealed unique +9.5 site activity to mediate GATA factor-dependent chromatin structural transitions. Loss-of-function analyses provided evidence for a mechanism in which a mediator of long-range transcriptional control [LIM domain binding 1 (LDB1)] and a chromatin remodeler [Brahma related gene 1 (BRG1)] synergize through the +9.5 site, conferring expression of GATA-2, which is known to promote the genesis and survival of hematopoietic stem cells.cis element | HSCs W hereas proximal promoter sequences assemble the basal transcriptional machinery and RNA polymerase, distant cis-regulatory elements often confer tissue-specific or contextdependent transcriptional regulation. Enhancer elements reside many kilobases upstream or downstream of a promoter or within introns, and extensive efforts have focused on elucidating "action-at-a-distance" mechanisms (1). Long-range transcriptional control involves physical interactions between proteins bound at distal regions and promoter sequences and higher order structural transitions, including subnuclear relocalization of target loci (2-4). Given the high frequency of long-range mechanisms at mammalian loci and the mutations that disrupt the function of such elements in pathological conditions, elucidating the underlying mechanisms in development,

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Section: Sequence Conservation and Transcription Factor Occupancy Do Notmentioning

confidence: 82%

Section: Significancementioning

confidence: 99%

Mechanism governing a stem cell-generating cis -regulatory element

Sanalkumar

Johnson

Gào

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…Conditional activation of ER-GATA-1 stably expressed in GATA-1-null G1E cells (G1E-ER-GATA-1 cells) induces differentiation, recapitulating a normal window of erythropoiesis (41). Treating G1E-ER-GATA-1 cells with increasing concentrations of ␤-estradiol or tamoxifen yields graded ER-GATA-1 occupancy at chromatin sites (13,21). ER-GATA-1 activation involves both increased ER-GATA-1 levels (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…At the Gata2 and ␣-globin loci, GATA-1 and GATA-2 have common and unique target sites. GATA-2 occupies sites 3.9, 2.8, and 1.8 kb upstream of the active Gata2 1S promoter (20,21). GATA-1-induced Gata2 repression involves ''GATA switches'' in which GATA-1 replaces GATA-2 at Ϫ3.9-and Ϫ2.8-kb regions.…”

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confidence: 99%

Differential sensitivities of transcription factor target genes underlie cell type-specific gene expression profiles

Johnson

Kim

Boyer

et al. 2007

Blood Cells, Molecules, and Diseases

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“…[28][29][30] GATA2 and GATA1 compete for occupancy of the same GATA-binding motifs in the Gata2 gene, and GATA2 enhances, but GATA1 represses, its transcription. 46,58,59 We previously demonstrated stable GATA1 expression (half-life Ͼ 6 hours) in erythroid leukemia cells, 31 in contrast to the rapid degradation of GATA2. GATA1 has neither an optimal motif for Cdk2 or Cdk4, nor a possible cyclin boxlike structure.…”

Section: Discussionmentioning

confidence: 99%

Cell-cycle–dependent oscillation of GATA2 expression in hematopoietic cells

Koga

Yamada

Abe

et al. 2007

Blood

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In vitro manipulation of hematopoietic stem cells (HSCs) is a key issue in both transplantation therapy and regenerative medicine, and thus new methods are required to achieve HSC expansion with self-renewal. GATA2 is a transcription factor controlling pool size of HSCs. Of interest, continuous overexpression of GATA2 does not induce HSC proliferation. In this report, we demonstrate that GATA2 expression, in leukemic and normal hematopoietic cells, oscillates during the cell cycle, such that expression is high in S phase but low in G 1 /S and M phase.GATA2 binding to target Bcl-X gene also oscillates in accordance with GATA2 expression. Using a green fluorescent protein (GFP)-GATA2 fusion protein, we demonstrate cell-cycle-specific activity of proteasome-dependent degradation of GATA2. Immunoprecipitation/immunoblotting analysis demonstrated phosphorylation of GATA2 at cyclin-dependent kinase (Cdk)-consensus motifs, S/T 0 P ؉1 , and interaction of GATA2 with Cdk2/ cyclin A2-, Cdk2/cyclin A2-, and Cdk4/ cyclin D1-phosphorylated GATA2 in vitro. Mutants in phosphorylation motifs exhibited altered expression profiles of GFP-GATA2 domain fusion proteins. These results indicate that GATA2 phosphorylation by Cdk/cyclin systems is responsible for the cell-cycle-dependent regulation of GATA2 expression, and suggest the possibility that a cell-cycle-specific "onoff" response of GATA2 expression may control hematopoietic-cell proliferation and survival. IntroductionHematopoietic stem cells (HSCs) give rise to the immunohematopoietic system throughout the lifetime of host animals. 1 HSCs have distinctive systems regulating their proliferation and differentiation. These systems maintain steady-state hematopoiesis, and respond to stresses, including infection, hypoxia, and blood loss. 1,2 Two types of cell division occur in HSCs: one type produces differentiated daughter cells to supply mature blood cells, while the other type reproduces HSCs (self-renewal division) to maintain life-long hematopoiesis. 1,2 Of interest, in the bone marrow reservoir of HSCs, more than 70% of bone marrow HSCs are quiescent (G 0 phase), maintaining high proliferative capacity and pluripotency. 3 In contrast, their quiescent mature progeny generally lack the capacity for growth and division.Directing the expansion of HSCs in vitro is an urgent problem that must be solved to meet the needs of transplantation therapy and to fulfill the promise of regenerative medicine. 4 HSC self-renewal and maturation divisions appear to be controlled, at least in part, by transcription factors (MEL/ELF4, 5 Gfi1 6,7 ), and also by cellcycle regulatory factors (cyclin-dependent kinase (Cdk) inhibitor, p21 Cip1/Waf1 , 8 p18 INK4C 9 phosphatase and tensin homologue (PTEN), 10 c-Myc, 11 and Myc antagonist, Mad 1 12 ). Analysis of mice deficient in Gfi1, 6,7 p21 Cip1/Waf1 , 8 or PTEN 10 revealed increased marrow HSC cycling and subsequent exhaustion of HSCs, and suggest possible cooperation between transcription factors and cell-cycle regulators.GATA2 is a member of th...

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Dynamic GATA Factor Interplay at a Multicomponent Regulatory Region of the GATA-2 Locus

Cited by 81 publications

References 51 publications

Mechanism governing a stem cell-generating cis -regulatory element

Mechanism governing a stem cell-generating cis -regulatory element

Differential sensitivities of transcription factor target genes underlie cell type-specific gene expression profiles

Cell-cycle–dependent oscillation of GATA2 expression in hematopoietic cells

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