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

Yan Li; Lei Wang; Jiawei Zhou; Fenge Li

doi:10.7717/peerj.2186

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

PeerJ. 2016 Jul 14:4:e2186. doi: 10.7717/peerj.2186. eCollection 2016.

Authors

Yan Li^#¹, Lei Wang^#¹, Jiawei Zhou¹, Fenge Li¹

Affiliation

¹ Key Laboratory of Pig Genetics and Breeding of Ministry of Agriculture & Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Huazhong Agricultural University, Wuhan, PR China.

^# Contributed equally.

Abstract

Klotho (KL), originally discovered as an aging suppressor, is a membrane protein that shares sequence similarity with the β-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 porcine KL gene. Deletion fragment analysis identified KL-D2 (-418 bp to -3 bp) as the porcine KL core 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 in KL binding 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 endogenous KL expression by RNA interference experiments. Our study indicates SNP MARC0022311 affects porcine KL expression by regulating its promoter activity via OCT-1.

Keywords: KL gene; MARC0022311; OCT-1; Pig.

Grants and funding

This work was supported financially by Key Projects in National Science R&T Program (2015BAD03B02, 2014BAD20B01), Hubei Science R&T Program (2014BBB008, 2014BBA194), and Fundamental Research Funds for the Central Universities. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.