GABP and PU.1 Compete for Binding, yet Cooperate to Increase CD18 (β2 Leukocyte Integrin) Transcription

Rosmarin, Alan G.; Caprio, David G.; Kirsch, David G.; Handa, Hiroshi; Simkevich, Carl P.

doi:10.1074/jbc.270.40.23627

Cited by 88 publications

(84 citation statements)

References 36 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The pPUBF⌬N-box is a pPUBF derivative with a deleted N-box/EBS sequence, and it has been previously described by our laboratory (Khurana et al, 1999). Expression vectors containing human GABP␣ and -␤1 subunits used were GABP␣ and GABP␤ (Rosmarin et al, 1995), and they were kindly gifted by Prof. Alan Rosmarin (Brown University). Expression vectors containing human ERF cDNA used were pSG5-ERF and GFP-ERF (Sgouras et al, 1995;Le Gallic et al, 1999), and they were kindly gifted by Prof. George Mavrothalassitis (IMBB, Greece).…”

Section: Constructsmentioning

confidence: 99%

“…To demonstrate the effect of ERF on utrophin-A promoter activity, we used the pPUBF utrophin-A luciferase reporter construct containing the entire human utrophin promoter (Dennis et al, 1996), as well as the pUBF⌬N-box reporter construct that contains a deletion mutation of the N-box region (Khurana et al, 1999) (Figure 2Bi). Drosophila S2 cells were cotransfected with pPUBF and 1) expression constructs encoding GABP␣/␤ (Rosmarin et al, 1995) as a positive control or 2) ERF expression vector pSG5-ERF. In agreement with previous studies (Khurana et al, 1999;Gyrd-Hansen et al, 2002), GABP␣/␤ trans-activated the utrophin A promoter fourfold (407 Ϯ 39.2%) (Figure 2Bii).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Ets-2 Repressor Factor Silences Extrasynaptic Utrophin by N-Box–mediated Repression in Skeletal Muscle

Perkins

Basu

Budak

et al. 2007

MBoC

View full text Add to dashboard Cite

Utrophin is the autosomal homologue of dystrophin, the protein product of the Duchenne's muscular dystrophy (DMD) locus. Utrophin expression is temporally and spatially regulated being developmentally down-regulated perinatally and enriched at neuromuscular junctions (NMJs) in adult muscle. Synaptic localization of utrophin occurs in part by heregulin-mediated extracellular signal-regulated kinase (ERK)-phosphorylation, leading to binding of GABP␣/␤ to the N-box/EBS and activation of the major utrophin promoter-A expressed in myofibers. However, molecular mechanisms contributing to concurrent extrasynaptic silencing that must occur to achieve NMJ localization are unknown. We demonstrate that the Ets-2 repressor factor (ERF) represses extrasynaptic utrophin-A in muscle. Gel shift and chromatin immunoprecipitation studies demonstrated physical association of ERF with the utrophin-A promoter N-box/EBS site. ERF overexpression repressed utrophin-A promoter activity; conversely, small interfering RNA-mediated ERF knockdown enhanced promoter activity as well as endogenous utrophin mRNA levels in cultured muscle cells in vitro. Laser-capture microscopy of tibialis anterior NMJ and extrasynaptic transcriptomes and gene transfer studies provide spatial and direct evidence, respectively, for ERF-mediated utrophin repression in vivo. Together, these studies suggest "repressing repressors" as a potential strategy for achieving utrophin up-regulation in DMD, and they provide a model for utrophin-A regulation in muscle. INTRODUCTIONDuchenne's muscular dystrophy (DMD) is a fatal neuromuscular disease caused by gene mutations leading to qualitative or quantitative disturbances in dystrophin expression (Hoffman et al., 1987). Dystrophin-related protein or utrophin is considered the autosomal homologue of dystrophin, because it shares extensive sequence homology as well as its size and abundance in muscle (Love et al., 1989;Khurana et al., 1990;Tinsley et al., 1992). Utrophin and dystrophin also share functional properties such as the ability to associate with the dystroglycan complex and bind F-actin (Matsumura et al., 1992;Winder and Kendrick, 1995;Rybakova et al., 2002). Direct evidence for the ability of utrophin to functionally substitute comes from experiments demonstrating that utrophin overexpression driven either by transgenic means Rafael et al., 1998;Tinsley et al., 1998;Fisher et al., 2001) or viral vectors (Gilbert et al., 1999;Wakefield et al., 2000;Cerletti et al., 2003) can rescue dystrophin-deficient muscle. Despite these functional similarities, dystrophin and utrophin exhibit distinct localization in normal adult tissues.Perinatally, utrophin is expressed throughout the sarcolemma, however, its expression declines as that of dystrophin increases (Khurana et al., 1991;Clerk et al., 1993), leading to its spatially restricted expression at neuromuscular and myotendinous junctions (NMJs and MTJs, respectively) of adult myofibers. These features and relevance toward a therapy for DMD provide a powerful impetus for b...

show abstract

Section: Constructsmentioning

confidence: 99%

mentioning

confidence: 99%

Ets-2 Repressor Factor Silences Extrasynaptic Utrophin by N-Box–mediated Repression in Skeletal Muscle

Perkins

Basu

Budak

et al. 2007

MBoC

View full text Add to dashboard Cite

show abstract

“…During the proliferation and differentiation of multipotential lymphoid-myeloid progenitors, PU.1 is expressed at all stages of granulocyte development, both during the early onset of expression of the GM-CSF and G-CSF receptors in myeloid progenitors (27)(28)(29)(30), and during the more terminal differentiation steps that include expression of the CD11b, CD18, and Fc␥RI receptors (31)(32)(33)(34)(35). PU.1 expression increases gradually through the myelocytic stages of hematopoietic development and then remains relatively constant through terminal differentiation (26,36).…”

Section: Family (C/ebp␣mentioning

confidence: 99%

Novel Combinatorial Interactions of GATA-1, PU.1, and C/EBPε Isoforms Regulate Transcription of the Gene Encoding Eosinophil Granule Major Basic Protein

Stankiewicz

Yang

et al. 2002

Journal of Biological Chemistry

106

139

View full text Add to dashboard Cite

GATA-1 and the ets factor PU.1 have been reported to functionally antagonize one another in the regulation of erythroid versus myeloid gene transcription and development. The CCAAT enhancer binding protein ⑀ (C/EBP⑀) is expressed as multiple isoforms and has been shown to be essential to myeloid (granulocyte) terminal differentiation. We have defined a novel synergistic, as opposed to antagonistic, combinatorial interaction between GATA-1 and PU.1, and a unique repressor role for certain C/EBP⑀ isoforms in the transcriptional regulation of a model eosinophil granulocyte gene, the major basic protein (MBP). The eosinophil-specific P2 promoter of the MBP gene contains GATA-1, C/EBP, and PU.1 consensus sites that bind these factors in nuclear extracts of the eosinophil myelocyte cell line, AML14.3D10. The promoter is transactivated by GATA-1 alone but is synergistically transactivated by low levels of PU.1 in the context of optimal levels of GATA-1. The C/EBP⑀ 27 isoform strongly represses GATA-1 activity and completely blocks GATA-1/PU.1 synergy. In vitro mutational analyses of the MBP-P2 promoter showed that both the GATA-1/PU.1 synergy, and repressor activity of C/EBP⑀ 27 are mediated via protein-protein interactions through the C/EBP and/or GATA-binding sites but not the PU.1 sites. Co-immunoprecipitations using lysates of AML14.3D10 eosinophils show that both C/EBP⑀ 32/30 and ⑀ 27 physically interact in vivo with PU.1 and GATA-1, demonstrating functional interactions among these factors in eosinophil progenitors. Our findings identify novel combinatorial protein-protein interactions for GATA-1, PU

show abstract

“…In addition to Sp1, the CD11b and c-fes promoters are regulated by PU.1 (28,31), and the CD18 promoter is regulated by PU.1 and GABP (14,32,33). Sp1 and GABP cooperatively activate the CD18 promoter in Schneider cells, the cytochrome c oxidase subunit IV promoter in NIH 3T3 or COS cells, and the folate-binding protein promoter in NIH 3T3 cells (34,35).…”

Section: Fig 4 Enhancer Segment C1 Binds Sp1 and Ets Factorsmentioning

confidence: 99%

An Enhancer Located between the Neutrophil Elastase and Proteinase 3 Promoters Is Activated by Sp1 and an Ets Factor

Nuchprayoon

Shang

Simkevich

et al. 1999

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

The adjacent neutrophil elastase, proteinase 3, and azurocidin genes encode serine proteases expressed specifically in immature myeloid cells. Subclones of a 17-kilobase (kb) murine neutrophil elastase genomic clone were assessed for their ability to stimulate the neutrophil elastase promoter in 32D cl3 myeloid cells. Region ؊9.3 to ؊7.3 kb stimulated transcription 7-fold, whereas other genomic segments were inactive. This enhancer is located in the second intron of the proteinase-3 gene and so may regulate more than one gene in the myeloid protease cluster. Deletional analysis of the enhancer identified several segments which activated the neutrophil elastase and thymidine kinase promoters 3-6-fold. The most active segment was a 220-base pair region centered at ؊8.6 kb, which activated transcription 31-fold. This segment contains an Sp1 consensus site, which bound Sp1, flanked by two Ets family consensus sequences, which bound PU.1, GABP, and an Ets factor present in myeloid cell extracts. Mutation of the Sp1-binding site reduced enhancer activity 8-fold in 32D cl3 cells, and mutation of either or both Ets-binding sites reduced activity 3-4-fold. Sp1 activated the distal enhancer 5-fold, GABP 3-fold, and the combination 8-fold in Schneider cells.To identify transcriptional events that determine myeloid cell determination, we have been investigating the regulation of the neutrophil elastase (NE) 1 gene. NE is a microbicidal serine protease present in the primary granules of granulocytes and monocytes (1). In bone marrow, NE mRNA is present mainly in promyelocytes (2), and transcriptional regulation plays a key role in its lineage-specific expression (3). We isolated a 17-kb genomic murine NE clone and found that, like the human NE gene, the murine gene contains five exons and initiates transcription 30 bp downstream from a TATAA homology (4). We demonstrated that the 5Ј-flanking region of the NE gene contains a 60-bp proximal enhancer, located just upstream of the TATAA homology, which is strongly active in 32D cl3 myeloid cells differentiating in response to granulocyte-colony-stimulating factor (G-CSF), but only weakly active in uninduced 32D cl3 cells or in fibroblasts (4). This proximal enhancer is activated cooperatively by C/EBP, c-Myb, CBF, and Ets family members such as PU.1 or GABP (4 -6). The NE gene is located just downstream of the proteinase-3 (PR-3) gene, which in turn is located just downstream of the azurocidin gene (7). These three serine protease genes are expressed specifically and coordinately in immature myeloid cells (7), and the promoter of each contains conserved binding sites for C/EBP, c-Myb, and PU.1 (7,8).To determine whether the murine NE gene contains additional regulatory regions, we assessed the activity of NE genomic subclones functionally after transient transfection into 32D cl3 cells. A 2-kb NE genomic region extending from Ϫ7.3 to Ϫ9.3 kb stimulated the NE promoter 7-fold, and stimulated the herpes TK promoter as well, whereas none of the other subclones, from Ϫ7.3 to ϩ7.5 kb...

show abstract

GABP and PU.1 Compete for Binding, yet Cooperate to Increase CD18 (β2 Leukocyte Integrin) Transcription

Cited by 88 publications

References 36 publications

Ets-2 Repressor Factor Silences Extrasynaptic Utrophin by N-Box–mediated Repression in Skeletal Muscle

Ets-2 Repressor Factor Silences Extrasynaptic Utrophin by N-Box–mediated Repression in Skeletal Muscle

Novel Combinatorial Interactions of GATA-1, PU.1, and C/EBPε Isoforms Regulate Transcription of the Gene Encoding Eosinophil Granule Major Basic Protein

An Enhancer Located between the Neutrophil Elastase and Proteinase 3 Promoters Is Activated by Sp1 and an Ets Factor

Contact Info

Product

Resources

About