scholarly journals Primary characterization and basal promoter activity of two hexamerin genes of Musca domestica

2004 ◽  
Vol 4 (2) ◽  
pp. 1-10 ◽  
Author(s):  
C. K. Moreira ◽  
Mde L. Capurro ◽  
M. Walter ◽  
E. Pavlova ◽  
H. Biessmann ◽  
...  
Keyword(s):  
Musca Domestica ◽  
Promoter Activity ◽  
Basal Promoter Activity

scholarly journals Primary characterization and basal promoter activity of two hexamerin genes of Musca domestica

2004 ◽  
Vol 4 (1) ◽  
Author(s):  
C.K. Moreira ◽  
M. de L. Capurro ◽  
M. Walter ◽  
E. Pavlova ◽  
H. Biessmann ◽  
...  
Keyword(s):  
Musca Domestica ◽  
Promoter Activity ◽  
Basal Promoter Activity

scholarly journals Regulation of basal promoter activity of the human thiamine pyrophosphate transporter SLC44A4 in human intestinal epithelial cells

2015 ◽  
Vol 308 (9) ◽  
pp. C750-C757 ◽  
Author(s):  
Svetlana M. Nabokina ◽  
Mel Brendan Ramos ◽  
Judith E. Valle ◽  
Hamid M. Said
Keyword(s):  
Gene Expression ◽  
Transcription Factors ◽  
Epithelial Cells ◽  
Promoter Activity ◽  
Minimal Promoter ◽  
Thiamine Pyrophosphate ◽  
Luciferase Reporter ◽  
Uptake System ◽  
Colonic Epithelial Cells ◽  
Basal Promoter Activity

Microbiota of the large intestine synthesize considerable amount of vitamin B1 in the form of thiamine pyrophosphate (TPP). There is a specific high-affinity regulated carrier-mediated uptake system for TPP in human colonocytes (product of the SLC44A4 gene). The mechanisms of regulation of SLC44A4 gene expression are currently unknown. In this study, we characterized the SLC44A4 minimal promoter region and identified transcription factors important for basal promoter activity in colonic epithelial cells. The 5′-regulatory region of the SLC44A4 gene (1,022 bp) was cloned and showed promoter activity upon transient transfection into human colonic epithelial NCM460 cells. With the use of a series of 5′- and 3′-deletion luciferase reporter constructs, the minimal genomic region that required basal transcription of the SLC44A4 gene expression was mapped between nucleotides −178 and +88 (using the distal transcriptional start site as +1). Mutational analysis performed on putative cis-regulatory elements established the involvement of ETS/ELF3 [E26 transformation-specific sequence (ETS) proteins], cAMP-responsive element (CRE), and SP1/GC-box sequence motifs in basal SLC44A4 promoter activity. By means of EMSA, binding of ELF3 and CRE-binding protein-1 (CREB-1) transcription factors to the SLC44A4 minimal promoter was shown. Contribution of CREB into SLC44A4 promoter activity was confirmed using NCM460 cells overexpressing CREB. We also found high expression of ELF3 and CREB-1 in colonic (NCM460) compared with noncolonic (ARPE19) cells, suggesting their possible contribution to colon-specific pattern of SLC44A4 expression. This study represents the first characterization of the SLC44A4 promoter and reports the importance of both ELF3 and CREB-1 transcription factors in the maintenance of basal promoter activity in colonic epithelial cells.

scholarly journals Rat gene 33: analysis of its structure, messenger RNA and basal promoter activity

1989 ◽  
Vol 17 (16) ◽  
pp. 6651-6667 ◽  
Author(s):  
Nancy B. Chrapkiewicz ◽  
Charles M. Davis ◽  
David T.-W. Chu ◽  
Catherine M. Caldwell ◽  
Daryl K. Granner
Keyword(s):  
Messenger Rna ◽  
Promoter Activity ◽  
Basal Promoter Activity

In vivo analyses of constitutive and regulated promoters in halophilic archaea

Microbiology ◽  
2005 ◽  
Vol 151 (1) ◽  
pp. 25-33 ◽  
Author(s):  
Dagmar Gregor ◽  
Felicitas Pfeifer
Keyword(s):  
Growth Phase ◽  
Promoter Activity ◽  
Halophilic Archaea ◽  
Stationary Growth Phase ◽  
Halobacterium Salinarum ◽  
Exponential Growth Phase ◽  
Haloferax Mediterranei ◽  
Basal Promoter Activity ◽  
Transcriptional Activator Protein

The two gvpA promoters PcA and PpA of Halobacterium salinarum, and the PmcA promoter of Haloferax mediterranei were investigated with respect to growth-phase-dependent expression and regulation in Haloferax volcanii transformants using the bgaH reading frame encoding BgaH, an enzyme with β-galactosidase activity, as reporter. For comparison, the Pfdx promoter of the ferredoxin gene of Hbt. salinarum and the PbgaH promoter of Haloferax lucentense (formerly Haloferax alicantei) were analysed. Pfdx , driving the expression of a house-keeping gene, was highly active during the exponential growth phase, whereas PbgaH and the three gvpA promoters yielded the largest activities during the stationary growth phase. Compared to Pfdx , the basal promoter activities of PpA and PmcA were rather low, and larger activities were only detected in the presence of the endogenous transcriptional activator protein GvpE. The PcA promoter does not yield a detectable basal promoter activity and is only active in the presence of the homologous cGvpE. To investigate whether the PcA -TATA box and the BRE element were the reason for the lack of the basal PcA activity, these elements and also sequences further upstream were substituted with the respective sequences of the stronger PpA promoter and investigated in Hfx. volcanii transformants. All these promoter chimera did not yield a detectable basal promoter activity. However, whenever the PpA -BRE element was substituted for the PcA -BRE, an enhanced cGvpE-mediated activation was observed. The promoter chimeras harbouring PpA -BRE plus 5 (or more) bp further upstream also gained activation by the heterologous pGvpE and mcGvpE proteins. The sequence required for the GvpE-mediated activation was determined by a 4 bp scanning mutagenesis with the 45 bp region upstream of PmcA -BRE. None of these alterations influenced the basal promoter activity, but the sequence TGAAACGG-n4-TGAACCAA was important for the GvpE-mediated activation of PmcA .

scholarly journals Adrenocorticotropin/3′,5′-Cyclic AMP-Mediated Transcription of the Scavenger akr1-b7 Gene in Adrenocortical Cells Is Dependent on Three Functionally Distinct Steroidogenic Factor-1-Responsive Elements

Endocrinology ◽  
2004 ◽  
Vol 145 (2) ◽  
pp. 508-518 ◽  
Author(s):  
Pierre Val ◽  
Christelle Aigueperse ◽  
Bruno Ragazzon ◽  
Georges Veyssière ◽  
Anne-Marie Lefrançois-Martinez ◽  
...  
Keyword(s):  
Adrenal Cortex ◽  
Promoter Activity ◽  
Specific Factor ◽  
Zona Fasciculata ◽  
Adrenocortical Cells ◽  
Steroidogenic Factor 1 ◽  
Basal Promoter Activity ◽  
Nuclear Extracts ◽  
Specific Complex ◽  
Drive Gene

Abstract The akr1-b7 gene encodes a scavenger enzyme expressed in steroidogenic glands under pituitary control. In the zona fasciculata of the adrenal cortex where its expression is controlled by ACTH, AKR1-B7 detoxifies isocaproaldehyde produced during the first step of steroidogenesis. Three steroidogenic factor-1 (SF-1)-responsive elements (SFREs) are contained within the −510/+41 promoter region, which was previously demonstrated to drive gene expression in transgenic mice adrenal cortex. All these sequences bind at least SF-1 in Y1 adrenocortical cell nuclear extracts and can be activated by overexpression of this factor in HeLa cells. However, the three SFREs show distinct properties regarding akr1-b7 promoter activity in Y1 cells. Whereas the proximal −102 SFRE supports basal promoter activity, the −458 bona fide SFRE is essential for both basal promoter activity and cAMP responsiveness, although it is unresponsive to cAMP when isolated from its promoter context. This suggests that SF-1 is not a cAMP-responsive factor per se. The neighboring SFRE at −503 is a palindromic sequence that binds monomeric and heteromeric SF-1 as well as an adrenal-specific complex. Using MA-10 Leydig cells and Y1–10r9 mutant cells, we provide evidence that its activity in adrenocortical cells depends on the binding of the adrenal-specific factor, which is required for basal and cAMP-induced promoter activity. Furthermore, the −503 site has intrinsic cAMP-sensing ability in Y1 cells, which is correlated with increased adrenal-specific complex binding. Collectively, our results suggest that cAMP responsiveness of the akr1-b7 promoter is achieved through cooperation between the adrenal-specific factor bound to the −503 site and SF-1 bound to the −458 site.

scholarly journals Localization of sequences for the basal and insulin-like growth factor-I inducible activity of the fatty acid synthase promoter in 3T3-L1 fibroblasts

1994 ◽  
Vol 298 (3) ◽  
pp. 575-578 ◽  
Author(s):  
S Misra ◽  
K Sakamoto ◽  
N Moustaïd ◽  
H S Sul
Keyword(s):  
Fatty Acid ◽  
Growth Factor ◽  
Promoter Activity ◽  
Fatty Acid Synthase ◽  
Luciferase Activity ◽  
Maximal Effect ◽  
Insulin Like Growth Factor ◽  
Factor I ◽  
Basal Promoter Activity ◽  
Igf I

Fatty acid synthase (FAS) plays a central role in fatty acid synthesis and its expression is under nutritional and hormonal control. We have investigated insulin-like growth factor-I (IGF-I) regulation of FAS by transfecting into 3T3-L1 fibroblasts chimeric genes comprising the 5′-flanking region of the FAS gene linked to a luciferase (LUC) reporter gene. First, the basal promoter activity of the 5′ serial deletions from nucleotides −318 to −19 of the FAS gene were compared. Deletions of the promoter sequences from −136 to −19 resulted in a step-wise decrease in the promoter activity, with the −67 LUC and −19 LUC plasmids retaining 40% and 16% of the luciferase activity of −136 LUC. Regulatory sequences important for the FAS basal promoter activity in 3T3-L1 fibroblasts are, therefore, located within the −136 to −19 region. Treatment with 10 mM IGF-I also increased luciferase activity 1.8 +/- 0.2-, 1.8 +/- 0.3- and 2.5 +/- 0.1-fold in 3T3-L1 fibroblasts transiently transfected with −136 LUC, −110 LUC and −67 LUC plasmids, respectively. Deletion of sequences from −67 to −19 resulted in the loss of responsiveness to IGF-I. Physiological doses of insulin (10 nM), however, did not increase luciferase activity in 3T3-L1 fibroblasts transfected with any of the above plasmids. Only upon treatment with pharmacological doses of insulin (1 microM), probably through IGF-I receptor, did luciferase activity increase 4.3 +/- 0.4-, 3.2 +/- 0.4- and 3.5 +/- 0.5-fold when transfected with −136 LUC, −110 LUC and −67 LUC plasmids, respectively; there was no increase with −19 LUC. The half-maximal effect of IGF-I on FAS promoter activity was observed at 3 nM and a maximal effect was reached at 10 nM. These results indicate that the increased promoter activities observed are probably mediated through the IGF-I receptor. Furthermore, sequences responsible for IGF-I regulation of the FAS gene are located within the proximal promoter between nucleotides −67 and −19 of the FAS gene.

Promoter cloning and characterization of the human programmed cell death protein 4 (pdcd4) gene: evidence for ZBP-89 and Sp-binding motifs as essential Pdcd4 regulators

2012 ◽  
Vol 32 (3) ◽  
pp. 281-297 ◽  
Author(s):  
Jörg Hendrik Leupold ◽  
Irfan Ahmed Asangani ◽  
Giridhar Mudduluru ◽  
Heike Allgayer
Keyword(s):  
Cell Death ◽  
Programmed Cell Death ◽  
Promoter Activity ◽  
Family Members ◽  
Tumour Suppressor ◽  
Solid Cancer ◽  
Basal Region ◽  
Mobility Shift ◽  
Basal Promoter Activity ◽  
Cell Death Protein

Pdcd4 (programmed cell death protein 4) is an important novel tumour suppressor inhibiting transformation, translation, invasion and intravasation, and its expression is down-regulated in several cancers. However, little is known about the transcriptional regulation and the promoter of this important tumour suppressor. So far the following is the first comprehensive study to describe the regulation of Pdcd4 transcription by ZBP-89 (zinc-finger-binding protein 89), besides characterizing the gene promoter. We identified the transcriptional start sites of the human pdcd4 promoter, a functional CCAAT-box, and the basal promoter region. Within this basal region, computer-based analysis revealed several potential binding sites for ZBPs, especially for Sp (specificity protein) family members and ZBP-89. We identified four Sp1/Sp3/Sp4-binding elements to be indispensable for basal promoter activity. However, overexpression of Sp1 and Sp3 was not sufficient to enhance Pdcd4 protein expression. Analysis in different solid cancer cell lines showed a significant correlation between pdcd4 and zbp-89 mRNA amounts. In contrast with Sp transcription factors, overexpression of ZBP-89 led to an enhanced expression of Pdcd4 mRNA and protein. Additionally, specific knockdown of ZBP-89 resulted in a decreased pdcd4 gene expression. Reporter gene analysis showed a significant up-regulation of basal promoter activity by co-transfection with ZBP-89, which could be abolished by mithramycin treatment. Predicted binding of ZBP-89 to the basal promoter was confirmed by EMSA (electrophoretic mobility-shift assay) data and supershift analysis for ZBP-89. Taken together, data for the first time implicate ZBP-89 as a regulator of Pdcd4 by binding to the basal promoter either alone or by interacting with Sp family members.

scholarly journals Mechanism for basal expression of rat mitochondrial branched-chain-2-oxo-acid dehydrogenase

1998 ◽  
Vol 334 (3) ◽  
pp. 713-722 ◽  
Author(s):  
Yi-Shuian HUANG ◽  
David T. CHUANG
Keyword(s):  
Dna Sequences ◽  
Binding Sites ◽  
Promoter Activity ◽  
Nuclear Protein ◽  
Luciferase Reporter ◽  
Kinase Gene ◽  
Acid Dehydrogenase ◽  
Basal Promoter Activity ◽  
Basal Expression ◽  
Branched Chain

The rat branched-chain-2-oxo-acid dehydrogenase (BCOD) kinase mRNA is transcribed from a TATA-less promoter that has GC-rich sequences and two putative Sp1 binding sites near the transcription start site. We demonstrated previously that the 5´ region of the kinase gene, base pairs -128 to +264, contained promoter activity when assayed using luciferase as a reporter (Huang and Chuang (1996) Biochem. J. 313, 603–609). To define DNA elements required for efficient expression of the kinase gene, nested deletion constructs of the above promoter region fused with a luciferase reporter gene were transfected into cultured H4IIE (hepatoma) and NRK-52E (kidney) cells. The results showed that the region between nucleotides -58 and +21 was indispensable for the kinase basal promoter activity. Methylation-interference and mutagenesis-promoter assays identified nucleotides -50 to -40 (ACAACTCCCA) as cis-acting DNA sequences that are required for nuclear protein binding and efficient promoter activity. Gel-supershift analysis with anti-Sp1 antibody suggested that the nuclear protein capable of binding to the -58 oligonucleotide (bp -58 to -34) was immunologically related to the Sp1 protein. The -58 oligonucleotide formed a DNA–protein complex with recombinant Sp1 protein with an affinity approximately ten-fold lower than that of the consensus Sp1 oligonucleotide. Co-transfection of the Sp1 expression plasmid and the -58 promoter construct into Drosophila Schneider cells revealed that Sp1 contributed to the kinase basal promoter activity by binding to the non-consensus site in the -58 region. Deletion of two consensus Sp1 binding sites (bases -150 to -140 and bases +29 to +38) in the kinase gene did not affect the basal promoter activity. Therefore binding of Sp1 or Sp1-like proteins to the above single non-consensus Sp1 sequence in the -58 region plays a major role of transactivating basal expression of the BCOD kinase.

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