scholarly journals Transcription factor organic cation transporter 1 (OCT-1) affects the expression of porcine Klotho (KL) gene

PeerJ ◽  
2016 ◽  
Vol 4 ◽  
pp. e2186 ◽  
Author(s):  
Yan Li ◽  
Lei Wang ◽  
Jiawei Zhou ◽  
Fenge Li
Keyword(s):  
Transcription Factor ◽  
Organic Cation ◽  
Core Promoter ◽  
Sequence Similarity ◽  
Organic Cation Transporter ◽  
Mobility Shift ◽  
Intron 1 ◽  
Organic Cation Transporter 1 ◽  
Immune Precipitation ◽  
Electrophoretic Mobility Shift Assays

Klotho (KL), originally discovered as an aging suppressor, is a membrane protein that shares sequence similarity with theβ-glucosidase enzymes. Recent reports showed Klotho might play a role in adipocyte maturation and systemic glucose metabolism. However, little is known about the transcription factors involved in regulating the expression of porcineKLgene. Deletion fragment analysis identified KL-D2 (−418 bp to −3 bp) as the porcineKLcore promoter. MARC0022311SNP (A or G) in KL intron 1 was detected in Landrace × DIV pigs using the Porcine SNP60 BeadChip. The pGL-D2-A and pGL-D2-G were constructed with KL-D2 and the intron fragment of different alleles and relative luciferase activity of pGL3-D2-G was significantly higher than that of pGL3-D2-A in the PK cells and ST cells. This was possibly the result of a change inKLbinding ability with transcription factor organic cation transporter 1 (OCT-1), which was confirmed using electrophoretic mobility shift assays (EMSA) and chromatin immune-precipitation (ChIP). Moreover, OCT-1 regulated endogenousKLexpression by RNA interference experiments. Our study indicates SNP MARC0022311 affects porcineKLexpression by regulating its promoter activity via OCT-1.

scholarly journals Transcription factor organic cation transporter 1 (OCT-1) affects the expression of porcine Klotho (KL) gene

2016 ◽  
Author(s):  
Yan Li ◽  
Lei Wang ◽  
Jiawei Zhou ◽  
Fenge Li
Keyword(s):  
Transcription Factor ◽  
Organic Cation ◽  
Core Promoter ◽  
Sequence Similarity ◽  
Organic Cation Transporter ◽  
Mobility Shift ◽  
Fragment Analysis ◽  
Intron 1 ◽  
Organic Cation Transporter 1 ◽  
Electrophoretic Mobility Shift Assays

Klotho (KL), originally discovered as an aging suppressor, was a membrane protein that shared sequence similarity with the β-glucosidase enzymes. Recent reports showed Klotho might have a role in adipocyte maturation and systemic glucose metabolism. However, little is known about the transcription factors involved in regulating the expression of porcine KL gene. Deletion fragment analysis identified KL-D2 (-418 bp to -3 bp) as the porcine KL core promoter. MARC0022311 in KL intron 1 appeared a polymorphism (A or G) in Landrace × DIV pigs, and relative luciferase activity of pGL3-D2-G was significantly higher than pGL3-D2-A. This was possibly the result of a change in KL binding ability with transcription factor organic cation transporter 1 (OCT-1), which was confirmed using electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP). Moreover, OCT-1 regulated endogenous KL expression by RNA interference. Our study indicates SNP MARC0022311 affects porcine KL expression by regulating its promoter activity via OCT-1.

scholarly journals Transcription factor organic cation transporter 1 (OCT-1) affects the expression of porcine Klotho (KL) gene

2016 ◽  
Author(s):  
Yan Li ◽  
Lei Wang ◽  
Jiawei Zhou ◽  
Fenge Li
Keyword(s):  
Transcription Factor ◽  
Organic Cation ◽  
Core Promoter ◽  
Sequence Similarity ◽  
Organic Cation Transporter ◽  
Mobility Shift ◽  
Fragment Analysis ◽  
Intron 1 ◽  
Organic Cation Transporter 1 ◽  
Electrophoretic Mobility Shift Assays

Klotho (KL), originally discovered as an aging suppressor, was a membrane protein that shared sequence similarity with the β-glucosidase enzymes. Recent reports showed Klotho might have a role in adipocyte maturation and systemic glucose metabolism. However, little is known about the transcription factors involved in regulating the expression of porcine KL gene. Deletion fragment analysis identified KL-D2 (-418 bp to -3 bp) as the porcine KL core promoter. MARC0022311 in KL intron 1 appeared a polymorphism (A or G) in Landrace × DIV pigs, and relative luciferase activity of pGL3-D2-G was significantly higher than pGL3-D2-A. This was possibly the result of a change in KL binding ability with transcription factor organic cation transporter 1 (OCT-1), which was confirmed using electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP). Moreover, OCT-1 regulated endogenous KL expression by RNA interference. Our study indicates SNP MARC0022311 affects porcine KL expression by regulating its promoter activity via OCT-1.

Regulation of basal core promoter activity of human organic cation transporter 1 (OCT1/SLC22A1)

2008 ◽  
Vol 295 (6) ◽  
pp. G1211-G1216 ◽  
Author(s):  
Moto Kajiwara ◽  
Tomohiro Terada ◽  
Jun-ichi Asaka ◽  
Masayo Aoki ◽  
Toshiya Katsura ◽  
...  
Keyword(s):  
Promoter Region ◽  
Promoter Activity ◽  
Organic Cation ◽  
Core Promoter ◽  
Organic Cation Transporter ◽  
Luciferase Assay ◽  
Basal Core Promoter ◽  
Human Liver Cell ◽  
E Box ◽  
Organic Cation Transporter 1

Human organic cation transporter 1 (OCT1/SLC22A1) plays important roles in the hepatic uptake of cationic drugs. The functional characteristics of this transporter have been well evaluated, but molecular information regarding transcriptional regulation is limited. In the present study, therefore, we examined the gene regulation of OCT1 gene focusing on basal core expression. An ∼2.5-kb fragment of the OCT1 promoter region was isolated, and promoter activity was measured by luciferase assay in the human liver cell lines Huh7 and HepG2. Deletion analysis suggested that the region spanning −141/−69 was essential for the basal core transcriptional activity and that this region contained the sequence of a cognate E-box (CACGTG). The E-box is known to be bound by the basal transcription factors, upstream stimulating factors (USFs), and the functional involvements of USF1 and USF2 were confirmed by a transactivation effect, a mutational analysis of the E-box, and an electrophoretic mobility shift assay. The transactivation effect of USFs on the OCT1 promoter was further stimulated by hepatocyte nuclear factor 4α, a liver-enriched transcription factor. There were no polymorphisms in the proximal promoter region (about 400 bp) of OCT1 gene ( n = 109). These findings indicated that both USF1 and USF2 bind to an E-box sequence located in the OCT1 core promoter region and are required for the basal gene expression of this transporter.

scholarly journals Impact of Promoter Polymorphisms on the Transcriptional Regulation of the Organic Cation Transporter OCT1 (SLC22A1)

2018 ◽  
Vol 8 (4) ◽  
pp. 42 ◽  
Author(s):  
Kristin Bokelmann ◽  
Jürgen Brockmöller ◽  
Mladen Tzvetkov
Keyword(s):  
Transcription Factor ◽  
Promoter Activity ◽  
Organic Cation ◽  
Specific Binding ◽  
Organic Cation Transporter ◽  
Luciferase Reporter ◽  
Potential Contribution ◽  
Luciferase Reporter Gene ◽  
Promoter Polymorphisms ◽  
Mobility Shift

The organic cation transporter 1 (OCT1, SLC22A1) is strongly expressed in the human liver and facilitates the hepatic uptake of drugs such as morphine, metformin, tropisetron, sumatriptan and fenoterol and of endogenous substances such as thiamine. OCT1 expression is inter-individually highly variable. Here, we analyzed SNPs in the OCT1 promoter concerning their potential contribution to the variability in OCT1 expression. Using electrophoretic mobility shift and luciferase reporter gene assays in HepG2, Hep3B, and Huh7 cell lines, we identified the SNPs −1795G>A (rs6935207) and −201C>G (rs58812592) as having effects on transcription factor binding and/or promoter activity. The A-allele of the −1795G>A SNP showed allele-specific binding of the transcription factor NF-Y leading to 2.5-fold increased enhancer activity of the artificial SV40 promoter. However, the −1795G>A SNP showed no significant effects on the native OCT1 promoter activity. Furthermore, the −1795G>A SNP was not associated with the pharmacokinetics of metformin, fenoterol, sumatriptan and proguanil in healthy individuals or tropisetron efficacy in patients undergoing chemotherapy. Allele-dependent differences in USF1/2 binding and nearly total loss in OCT1 promoter activity were detected for the G-allele of −201C>G, but the SNP is apparently very rare. In conclusion, common OCT1 promoter SNPs have only minor effects on OCT1 expression.

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