Transcription Control in Eucaryotes-Enhancers and Promoters

The gene for murine erythropoietin (EPO) was isolated from a mouse genomic library with a human EPO cDNA probe. Nucleotide sequence analysis permitted the identification of the murine EPO coding sequence and the prediction of the encoded amino acid sequence based on sequence conservation between the mouse and human EPO genes. Both the coding DNA and the amino acid sequences were 80% conserved between the two species. Transformation of COS-1 cells with a mammalian cell expression vector containing the murine EPO coding region resulted in secretion of murine EPO with biological activity on both murine and human erythroid progenitor cells. The transcription start site for the murine EPO gene in kidneys was determined. This permitted tentative identification of the transcription control region. The region included 140 base pairs upstream of the cap site which was over 90% conserved between the murine and human genes. Surprisingly, the first intron and much of the 5'- and 3'-untranslated sequences were also substantially conserved between the genes of the two species.

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Abstract 83: Modulation of Macrophage Function via Metabolism by Trypanosoma cruzi

Circulation Research ◽

10.1161/res.117.suppl_1.83 ◽

2015 ◽

Vol 117 (suppl_1) ◽

Author(s):

Sue-Jie Koo ◽

Nisha J Garg

Keyword(s):

Trypanosoma Cruzi ◽

Chronic Phase ◽

Etiologic Agent ◽

Acid Oxidation ◽

Peroxisome Proliferator ◽

Transcription Control ◽

Metabolic Gene Expression ◽

Anti Inflammatory ◽

Peroxisome Proliferator Activated Receptors ◽

The Impact

Chagas heart disease is an inflammatory cardiomyopathy which presents with mononuclear infiltrates in the interstitium and myocardial fibrosis in the chronic phase. Incomplete clearance by macrophages of the etiologic agent, Trypanosoma cruzi , is a significant cause of chronic disease development in approximately 30% of those serologically positive for the blood-borne parasite. The differential metabolic status, anaerobic glycolysis and mitochondria-dependent oxidative phosphorylation, are respectively associated with pro-inflammatory (M1) and anti-inflammatory (M2) functional activation of macrophages. Reactive oxygen species (ROS) have been shown to be an intracellular signal for glycolysis while peroxisome proliferator-activated receptors (PPARs) that enhance fatty acid oxidation provide transcription control of macrophage functional state. In our studies using diverse T. cruzi isolates, we showed that SylvioX10 (virulent), but not TCC (non-virulent), isolates are able to differentially control extracellular and intracellular ROS levels in macrophages. We found in macrophages infected with SylvioX10, the nuclear expression of PPAR-α was increased by 18 hours post-infection, and mitochondrial metabolic activity was similar to that of not-infected and M2 controls; which indicates anti-inflammatory function of macrophages, and therefore prohibiting T. cruzi clearance. In our ongoing studies, we are examining the impact of PPAR-α inhibitors in modulating the metabolic gene expression profile, functional phenotype and parasite survival in macrophages. Our data will provide the first indication that host macrophages have deficient pro-inflammatory capacity due to sub-optimal glucose oxidation, and enhancing the metabolism that supports T. cruzi clearance will provide a valuable basis for a strategy to arrest Chagas disease progression.

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TRES predicts transcription control in embryonic stem cells

Bioinformatics ◽

10.1093/bioinformatics/btu399 ◽

2014 ◽

Vol 30 (20) ◽

pp. 2983-2985 ◽

Cited By ~ 3

Author(s):

Christopher Pooley ◽

David Ruau ◽

Patrick Lombard ◽

Berthold Gottgens ◽

Anagha Joshi

Keyword(s):

Stem Cells ◽

Embryonic Stem Cells ◽

Embryonic Stem ◽

Transcription Control

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Functional properties of a Drosophila homolog of the E2F1 gene.

Molecular and Cellular Biology ◽

10.1128/mcb.14.3.1603 ◽

1994 ◽

Vol 14 (3) ◽

pp. 1603-1612 ◽

Cited By ~ 65

Author(s):

K Ohtani ◽

J R Nevins

Keyword(s):

Dna Binding ◽

Growth Cycle ◽

Glutathione S Transferase ◽

Transcription Control ◽

Cellular Transcription Factor ◽

C Terminus ◽

Considerable Homology ◽

Transcription Factor E2f ◽

Cellular Transcription

A variety of studies have now implicated the cellular transcription factor E2F as a key participant in transcription control during the cell growth cycle. Although the recent isolation of molecular clones encoding proteins that are components of the E2F activity (E2F1 and DP-1) provides an approach to defining the specific involvement of E2F in these events, definitive experiments remain difficult in the absence of appropriate genetic systems. We have now identified a Drosophila equivalent of E2F1 that we hope will allow an eventual genetic approach to the role of E2F in cellular regulatory events. A cDNA clone was isolated from a Drosophila cDNA library by using a probe containing sequence from the E2F1 DNA binding domain. The sequence of the clone, which we term drosE2F1, demonstrates considerable homology to the human E2F1 sequence, with over 65% identity in the DNA binding region and 50% identity in the region of E2F1 known to interact with the retinoblastoma gene product. A glutathione S-transferase-drosE2F1 fusion protein was capable of binding specifically to an E2F recognition site, and transfection assays demonstrated that the drosE2F1 product was capable of transcription activation, dependent on functional E2F sites as well as sequences within the C terminus of the protein. Finally, we have also identified E2F recognition sequences within the promoter of the Drosophila DNA polymerase alpha gene, and we demonstrate that the drosE2F1 product activates transcription of a test gene under the control of this promoter. We conclude that the drosE2F1 cDNA encodes an activity with extensive structural and functional similarity to the human E2F1 protein.

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