The Hematopoietic Transcription Factor PU.1 Represses Gelatinase A Transcription in Glomerular Mesangial Cells

Harendza, Sigrid; Lovett, David H.; Stahl, Rolf A.K.

doi:10.1074/jbc.m001322200

Cited by 17 publications

(16 citation statements)

References 41 publications

(17 reference statements)

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“…Among the known Ets family members, approximate sizes of 65 kD have been reported for Elf-1 and PEA3. Another member of the Ets protein family, PU.1, with a calculated molecular weight of 30.7 kD has recently been identified in MC as negative regulator of gelatinase A transcription that acts via an upstream silencer element (11). Ets transcription factors with molecular sizes of 15 kD have not been described, suggesting that this band constitutes a proteolytic cleavage product.…”

Section: Discussionmentioning

confidence: 99%

“…Reverse transcription was achieved by addition of AMV reverse transcriptase (Promega, Madison, WI) in a reaction buffer containing 4 l (40 U/l) of RNase inhibitor (Roche Molecular Biochemicals, Summerville, NJ), 0.5 g of oligo d(T) [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18] , 0.5 nM dNTPs, and incubation for 2 h at 42°C and for 5 min at 95°C. Subsequently, cDNAs were precipitated and PCR were performed by addition of 2 U Taq polymerase (Life Technologies-BRL, Grand Island, NY) in reaction buffer (200 mM TrisHCl [pH 8.3], 0.5 M KCl, 1.5 mM MgCl 2 ), 0.25 pmol dNTPs, 25 pmol primers (specific Ets-1 primers for full-length rat ets-1 transcripts were: 5'-ATGAAGGCGGCCGTC-GATCT-3' and 5'-TTACTCATCAGCATCCGGCTTCA-3').…”

Section: Reverse Transcription Pcrmentioning

confidence: 99%

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Transcription Factor Ets-1 Regulates Gelatinase A Gene Expression in Mesangial Cells

Reisdorff¹,

En‐Nia²,

Stefanidis³

et al. 2002

Journal of the American Society of Nephrology

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Abstract. Ets transcription factors are involved in cell growth and angiogenesis. Ets-1 targets include members of the matrix metalloproteinase superfamily. In inflammatory glomerular diseases, the patterns and regulation of Ets expression have not been fully characterized. In the present study, nuclear binding activities to the consensus Ets-1/PEA3 motif were detected in mesangial cells (MC), and the Ets-1 protein was positively identified by Western blotting, reverse transcription PCR (RT-PCR), and DNA-binding studies. The 5' flanking regions of the human and rat gelatinase A genes contain clusters of potential Ets-1 binding motifs, one of which is evolutionarily conserved. Using a series of 5' deletion reporter constructs of the rat gelatinase A gene and an Ets-1 expression plasmid, a concentration-dependent threefold trans-activation of gene expression mapped to the conserved Ets-1 binding motif at Ϫ1004/Ϫ1053 bps, designated responsive element-2 (RE-2). The RE-2 was operative within the context of the homologous gelatinase A promoter but not with a heterologous simian virus 40 promoter. Specific Ets-1 binding to this sequence was demonstrated by DNA-binding studies. Transient expression of an Ets-1 expression plasmid increased gelatinase A protein expression. Our findings identify an additional matrix metalloproteinase family member, gelatinase A, as an Ets-1 responsive gene in MC that may play a role in the high level expression of this enzyme in inflammatory glomerular diseases.Mesangial cell (MC) proliferation, increased matrix turnover, and accumulation of interstitial collagens are common findings in most forms of progressive glomerular diseases (1). These changes generally continue even after removal of the initial causative event, progressively resulting in destruction of the glomerular architecture with the final outcome of glomerular scarring (1). Recent findings have elucidated a pivotal role for the "activated MC" phenotype (2). In contrast to the quiescent MC found within normal glomeruli, activated MC predominantly secrete interstitial type I and III collagens. Concurrently, high-level expression of matrix metalloproteinase-2 (MMP-2, 72-kD type IV collagenase, gelatinase A) is found (3,4). Gelatinase A belongs to the matrix metalloproteinase gene superfamily, all of which are secreted as latent proenzymes with subsequent activation by proteolysis in the extracellular space. The enzymatic activity of the MMP is dependent on the presence of zinc and divalent cations (5), and these enzymes exhibit broad substrate specificity (6). In addition to the role of gelatinase A in glomerular extracellular matrix turnover, the enzyme has a direct influence on the MC phenotype. Turck et al. (7) have shown that targeted deletion of gelatinase A in MC results in an exit from the cell cycle and development of a phenotype resembling quiescent MC in vivo. Addition of exogenous gelatinase A, but not gelatinase B, promoted MC proliferation with reacquisition of the prosclerotic phenotype (7). Concordant gelatinas...

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Section: Discussionmentioning

confidence: 99%

Section: Reverse Transcription Pcrmentioning

confidence: 99%

Transcription Factor Ets-1 Regulates Gelatinase A Gene Expression in Mesangial Cells

Reisdorff¹,

En‐Nia²,

Stefanidis³

et al. 2002

Journal of the American Society of Nephrology

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“…Negative regulatory function of PU.1 has been described in the regulation of other genes. 26,27 Whether repression of IL-10 expression is exerted by PU.1 alone or by interactions with other transcription factors remains unclear. Interaction of PU.1 with other transcription factors is under investigation.…”

Section: Differential Regulation Of Il-10mentioning

confidence: 99%

Differential regulation of interleukin-10 production by genetic and environmental factors – a twin study

et al. 2002

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Interleukin-10 (IL-10) has a critical role in the regulation of immune responses. The relative contribution of genetic and environmental factors to IL-10 production is under debate. We performed a twin study in 246 monozygotic and dizygotic twins to assess the heritability of IL-10 production after LPS stimulation in whole blood. In addition, the influence of promoter single nucleotide polymorphisms (À1082, À819 and À592) on transcriptional activity and their binding to nuclear factors was studied in luciferase reporter gene and electrophoretic mobility shift assays. IL-10 production showed a genetic determination with a heritability of 0.5. Decreasing body mass index (BMI), smoking and female gender lead to decreased IL-10 production. In monocytes, the À1082A allele showed higher binding affinity to the transcription factor PU.1 resulting in decreased transcriptional activity of À1082A promoter haplotypes. Genetic determination of IL-10 secretion is probably lower than that previously reported. Fifty percent of the observed variability explained by genetic factors. Female individuals produce less IL-10 than male subjects. Environmental factors like smoking and decreasing BMI exert suppressing effects on IL-10 production. Although the À1082A allele shows higher binding affinity to the PU.1 transcription factor and lower transcriptional activity, this polymorphism probably explains only a small fraction of the observed heritability.

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“…In the promoter region of the rat MMP-2 gene, four regulatory cis-acting elements, at Ϫ1854/Ϫ1849 (PU.1-binding site), 13) Ϫ1394/Ϫ1385 (AP-1-binding site), 12) Ϫ1322/Ϫ1282 (designated RE-1, YB-1 · AP-2 · p53 ternary complex-binding site) 10) and Ϫ1021/Ϫ972 (designated RE-2, Ets-1-binding site), 14) have so far been identified. Next, it was examined whether these four regulatory elements identified in the 5Ј-flanking region of the rat MMP-2 gene are also conserved in the orthologous region of the mouse MMP-2 gene.…”

Section: Comparison Of the 5-flanking Regions Of The Mouse And Rat MMmentioning

confidence: 99%

“…13) In addition to the PU.1 binding element, AP-1 binding element and RE-1, a fourth region at positions Ϫ1021 to Ϫ972, designated RE-2, was identified as an Ets-1-binding site in which rat MMP-2 transcription was increased through Ets-1 transcription factor. 14) We previously showed that extracellular matrix tenascin-X-deficient (TNX-/-) mice exhibit promotion of tumor invasion and metastasis due to increased levels of MMP-2 and MMP-9 activities.…”

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confidence: 99%

Transcription Regulatory Complex Including YB-1 Controls Expression of Mouse Matrix Metalloproteinase-2 Gene in NIH3T3 Cells

Matsumoto

Abiko

Ariga

2005

Biological & Pharmaceutical Bulletin

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During metastasis, invasive cells must transverse tissue barriers comprised by a variety of extracellular matrices (ECM). This process depends on the ability of tumor cells to degrade the surrounding matrices and then migrate through the matrix defects.1) Matrix metalloproteinase (MMP)-2 (type IV collagenase; gelatinase A) has been suggested to play a role in tumor metastasis.2) The activity of MMP-2 is regulated at three steps: gene expression by several transcription factors, proenzyme processing by tissue inhibitors of metalloproteinase-2 and membrane-type metalloproteinases, and inhibition of enzymatic activity by naturally occurring tissue inhibitors of metalloproteinases.3) Recently, the mechanisms of human MMP-2 gene expression in cancer cells have become clear. Qin et al. 4) demonstrated that several transcription factors, including Sp1, Sp3 and AP-2, participate in the control of constitutive MMP-2 gene expression. The same group also demonstrated that Brg-1 plays a role in the regulation of constitutive expression of the MMP-2 gene by recruitment of Sp1, AP-2 and Pol II to the MMP-2 promoter.5) Furthermore, p53 has been also shown to transcriptionally activate the expression of the human MMP-2 gene.6) Interestingly, it was reported that ATF3 interferes with human MMP-2 expression by antagonizing p53-dependent activation of the MMP-2 gene promoter. 7)On the other hand, four regulatory cis-elements for the gene expression of rat MMP-2 have been identified. Analyses of rat MMP-2 gene transcription have demonstrated that specific interaction of the nuclear factor YB-1, a member of the Y-box transcription factor family, with a 40-bp enhancer element, denoted as RE-1, located at nucleotide positions Ϫ1322 to Ϫ1282 relative to the translational start site is necessary for potent expression of the rat MMP-2 gene.8,9) The same authors have also shown that combinatorial interaction of AP-2 and p53 with YB-1 in RE-1 yields more increased MMP-2 expression.10) Another regulator of rat MMP-2 transcription, nm23-b, that acts by competitive interference with the transactivator YB-1, leading to suppression of MMP-2 transcription, has also been identified in RE-1.11) Moreover, under hypoxic conditions, MMP-2 transcription has been reported to be regulated in an AP-1-binding site located at positions Ϫ1394 to Ϫ1385 through JunB/Fra1 and JunB/FosB heterodimers.12) An additional silencer element in which PU.1 transcription factor binds was isolated in a further upstream region (positions Ϫ1854 to Ϫ1849) of the rat MMP-2 gene.13) In addition to the PU.1 binding element, AP-1 binding element and RE-1, a fourth region at positions Ϫ1021 to Ϫ972, designated RE-2, was identified as an Ets-1-binding site in which rat MMP-2 transcription was increased through Ets-1 transcription factor. 14)We previously showed that extracellular matrix tenascin-X-deficient (TNX-/-) mice exhibit promotion of tumor invasion and metastasis due to increased levels of MMP-2 and MMP-9 activities.15) It has also been revealed that the induction of MMP...

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The Hematopoietic Transcription Factor PU.1 Represses Gelatinase A Transcription in Glomerular Mesangial Cells

Cited by 17 publications

References 41 publications

Transcription Factor Ets-1 Regulates Gelatinase A Gene Expression in Mesangial Cells

Transcription Factor Ets-1 Regulates Gelatinase A Gene Expression in Mesangial Cells

Differential regulation of interleukin-10 production by genetic and environmental factors – a twin study

Transcription Regulatory Complex Including YB-1 Controls Expression of Mouse Matrix Metalloproteinase-2 Gene in NIH3T3 Cells

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