The Aldolase A Promoter in Proliferating Rat Thymocytes Is Regulated by a Cluster of SP1 Sites and a Distal Modulator

Hermfisse, Ulrich; Schäfer, Doris; Netzker, Roland; Brand, Karl

doi:10.1006/bbrc.1996.1284

Cited by 14 publications

(10 citation statements)

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“…A negative regulatory transcriptional element (reducing module), located upstream from exon M (clone Ϫ1066/Ϫ731), has been found to attenuate the proximal promoter (AH1)-driven transcription in the rat thymocyte aldolase A gene (21). However, this negative "reducing module" is located in a position completely different from that of the AldA-NRE.…”

Section: Discussionmentioning

confidence: 91%

Negative Regulation of the Mouse Aldolase A Gene

Lupo

Costanzo

Medugno

et al. 1997

Journal of Biological Chemistry

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The expression of aldolase A L-type mRNA is increased in growth-arrested mouse NIH3T3 cells and remarkably down-regulated in actively proliferating cells. Treatment of proliferating cells with cycloheximide abolished the down-regulation of L-type mRNA expression, thus indicating that a protein factor acts as repressor in proliferating cells. Transient transfection experiments in NIH3T3 cells showed that a negative regulatory cis-element (NRE) is involved in the modulation of the transcriptional activity of the distal L promoter. The repressor, which is a protein of ϳ97 kDa, binds the murine aldolase A NRE, revealing a much more intense DNA-protein complex in proliferating NIH3T3 cells than in serum-deprived cells. Mutations in the negative regulatory cis-element showed that the GArich motif is required for protein binding and silencer function. We conclude that the expression of L-type mRNA is modulated by the interaction between a cell cycle-dependent DNA-binding protein and the murine aldolase A NRE.Aldolase A gene expression is regulated by an intriguing mechanism that involves three alternative promoter regions (distal or pL, middle or pM, and proximal or pF). Transcription of multiple mRNA species (L, M, and F types), containing the same coding region but different untranslated 5Ј-ends, is driven by three autonomous promoters and splicing of alternative leader exons (L, M, and F) (1-3). In man, two of these mRNA species (L and F types) are ubiquitously expressed, whereas the third one (M type) is muscle-specific (4 -6).We have demonstrated that the human genomic regions upstream from exons F and L are able to drive autonomous transcription (7,8). We found that the basal transcriptional activity of pF is regulated by several binding sites for the ubiquitous factor Sp1 (7). Binding of nuclear trans-acting factors AP-1 and NF-1 in the Ϫ384/Ϫ262 region increases pF transcriptional activity (7, 9).The pL promoter is regulated by both positive and negative cis-elements (8). Within this promoter, we detected a negative regulatory element (hAldA-NRE) 1 and a protein factor from Hep3B cells that binds to this element. This complex downregulates to one-fifth the transcription of the distal promoter in several cell types and furthermore regulates transcription of other cellular genes (10).In rodent tissues, only two mRNAs, the ubiquitous AH type, corresponding to the human F type, and the muscle-specific type (M type), have been characterized (11,12). We found that a third species, which corresponds to human aldolase A L-type mRNA, is expressed and correctly processed in rodent cell lines and that its expression is increased during differentiation and is associated with cell-growth arrest (13).Here we report that the modulation of murine L-type mRNA expression is transcriptionally controlled and differentially regulated in proliferating and confluence-arrested cell populations. Within the murine distal promoter, we identified a negative regulatory element (mAldA-NRE), homologous to the human aldolase A silencer, that ...

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Section: Discussionmentioning

confidence: 91%

Negative Regulation of the Mouse Aldolase A Gene

Lupo

Costanzo

Medugno

et al. 1997

Journal of Biological Chemistry

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“…Sp1 plays a critical role in the transcription of a number of glycolytic enzymes (55,66,67). Thus, it is conceivable that Sp1 and O-GlcNAc function as glucose sensors that link the metabolic activities of a cell to its nutritional condition.…”

Section: Discussionmentioning

confidence: 99%

Down-regulation of Sp1 Activity through Modulation of O-Glycosylation by Treatment with a Low Glucose Mimetic, 2-Deoxyglucose

Kang

Cho

et al. 2003

Journal of Biological Chemistry

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2-Deoxyglucose (2-DG), a nonmetabolizable glucose analogue, blocks glycolysis at the phosphohexose isomerase step and has been frequently used as a glucose starvation mimetic in studies of a wide variety of physiological dysfuctions. However, the effect of 2-DG on protein glycosylation and related signal pathways has not been investigated in depth. In HeLa, an HPV18-positive cervical carcinoma line, 2-DG treatment downregulates human papillomavirus early gene transcription. This down-regulation was also achieved by low glucose supply or hypoxia, suggesting that this is a response commonly modulated by cellular glucose or energy level. We investigated how 2-DG and low glucose affect transcriptional activity. Human papillomavirus gene transcription was only marginally affected by the inhibition of ATP synthesis or the supplementation of pyruvate to 2-DG-treated cells, suggesting that poor ATP generation is involved only to a limited extent. 2-DG treatment also inhibited activation of p21 WAF1 promoter, which is controlled by p53 and/or Sp1. In a reporter assay using p21 WAF1 promoter constructs, 2-DG exerted a strong inhibitory effect on Sp1 activity. DNA binding activity of Sp1 in 2-DG-treated HeLa cells was intact, whereas it was severely impaired in cells incubated in a low glucose medium or in hypoxic condition. Unexpectedly, Sp1 was heavily modified with GlcNAc in 2-DG-treated cells, which is at least partially attributed to the inhibitory effect of 2-DG on N-acetyl-␤-D-glucosaminidase activity. Our results suggest that 2-DG, like low glucose or hypoxic condition, down-regulates Sp1 activity, but through hyper-GlcNAcylation instead of hypo-GlcNAcylation.

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“…Previous work investigating metabolic differences between resting and dividing thymocytes has revealed the importance of intracellular redox upon the growthspecific control of metabolic gene expression (Netzker et al, 1994;Hermfisse et al, 1996;Schafer et al, 1996;Brand and Hermfisse, 1997;Hamm-Kunzelmann et al, 1997;Brand et al, 2000). One possible mechanism by which cells regulate growth-specific gene expression in a redox-specific fashion is the relative abundance of reactive oxygen species (ROS).…”

Section: Cells and Hepg2 Cells Under Different Growth Conditionsmentioning

confidence: 99%

Cystathionine β‐synthase is coordinately regulated with proliferation through a redox‐sensitive mechanism in cultured human cells and Saccharomyces cerevisiae

Maclean

Jánošík

Kraus

et al. 2002

Journal Cellular Physiology

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Cystathionine beta-synthase (CBS) catalyzes the condensation of serine with homocysteine to form cystathionine and occupies a crucial regulatory position between the methionine cycle and the biosynthesis of cysteine by transsulfuration. Analysis of CBS activity under a variety of growth conditions indicated that CBS is coordinately regulated with proliferation in both yeast and human cells. In batch cultures of Saccharomyces cerevisiae, maximal CBS activities were observed in the exponential phase of cells grown on glucose, while growth-arrested cultures or those growing non-fermentatively on ethanol or glycerol had approximately 3-fold less activity. CBS activity assays and Western blotting indicated that growth-specific regulation of CBS is evolutionarily conserved in a range of human cell lines. CBS activity was found to be maximal during proliferation and was reduced two- to five-fold when cells became quiescent at confluence. In cultured HepG2 cells, the human CBS gene is induced by serum and basic fibroblast growth factor and is downregulated, but not abolished, by contact inhibition, serum-starvation, nutrient depletion, or the induction of differentiation. Consequently, for certain cell types, CBS may represent a novel marker of both differentiation and proliferation. The intracellular level of the CBS regulator compound, S-adenosylmethionine, was found to reflect the proliferation status of both yeast and human cells, and as such, constitutes an additional mechanism for proliferation-specific regulation of human CBS. Our data indicates that screening compounds for the ability to affect transsulfuration in cultured cell models must take proliferation status into account to avoid masking regulatory interactions that may be of significance in vivo.

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The Aldolase A Promoter in Proliferating Rat Thymocytes Is Regulated by a Cluster of SP1 Sites and a Distal Modulator

Cited by 14 publications

References 0 publications

Negative Regulation of the Mouse Aldolase A Gene

Negative Regulation of the Mouse Aldolase A Gene

Down-regulation of Sp1 Activity through Modulation of O-Glycosylation by Treatment with a Low Glucose Mimetic, 2-Deoxyglucose

Cystathionine β‐synthase is coordinately regulated with proliferation through a redox‐sensitive mechanism in cultured human cells and Saccharomyces cerevisiae

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