Transcription factor organic cation transporter 1 (OCT-1) affects the expression of porcine Klotho (KL) gene
- Published
- Accepted
- Copyright
- © 2016 Li et al.
- Licence
- This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ Preprints) and either DOI or URL of the article must be cited.
- Cite this article
- 2016. Transcription factor organic cation transporter 1 (OCT-1) affects the expression of porcine Klotho (KL) gene. PeerJ Preprints 4:e1800v1 https://doi.org/10.7287/peerj.preprints.1800v1
Abstract
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.
Author Comment
This is a submission to PeerJ for review.
Supplemental Information
Transcription factor binding site prediction of the procine KL intron 1 containing MARC0022311 (KL g.1474 A>G)
Quadrilateral frame indicated the substitutions and extra binding site of OCT-1. (A) Predicted by BIOBASE online software. (B) Predicted by TFserach online software.
Genotyping results of MARC0022311
(A) PK cells. (B) ST cells. MARC0022311 was marked in gray backgound.
OCT-1 binding sites in the porcine KL intron 1
(A) Frequency distribution of the predicted OCT-1 binding sites. X-axis indicated the length of the porcine KL intron 1 in bp. Y-axis was the frequency of the predicted OCT-1 binding sites. (B) ChIP analysis of three candidate OCT-1 binding sites (1395 bp to 1525 bp, 14322 bp to 14436 bp, 30970 bp to 31141 bp) in KL intron 1 in PK cells. Primers used for ChIP-PCR was shown in Table 1. Input and R were positive control, while IgG was the negative control.
