Functional analysis of haplotypes and promoter activity at the 5' region of the human GABRB3 gene and associations with schizophrenia.

Abstract

BACKGROUND:

This study investigated the effects of haplotypes T-G and C-A derived from NG_012836.1:g.4160T>C and NG_012836.1:g.4326G>A on protein expression levels in vitro and identified the functional sequence in the regulatory region of the GABRB3 gene linked to possible associations with schizophrenia.

METHODS:

Recombinant plasmids with haplotypes T-G and C-A and 10 recombinant vectors containing deletion fragments from the GABRB3 gene 5' regulatory region were transfected into HEK-293, SK-N-SH, and SH-SY5Y cells. The relative fluorescence intensity of the two haplotypes and different sequences was compared using a dual luciferase reporter assay system.

RESULTS:

The relative fluorescence intensity of haplotype C-A was significantly lower than that of T-G. We shortened the core promoter sequence of the GABRB3 gene 5' regulation region from -177 bp to -18 bp (ATG+1). We also found an expression suppression region from -1,735 bp to -1,638 bp and an enhanced regulatory region from -1,638 bp to -1,335 bp. Multiple inhibitory functional elements were identified in the region from -680 bp to -177 bp.

CONCLUSION:

We demonstrated that haplotype C-A might increase the risk of schizophrenia and found multiple regulatory regions that had an effect on GABRB3 receptor expression.

• Authors: Yi Liu, Mei Ding, Yong-Ping Liu, Xi-Cen Zhang, Jia-Xin Xing, Jin-Feng Xuan, Xi Xia, Jun Yao, Bao-Jie Wang
• Journal: Molecular genetics & genomic medicine (Mol Genet Genomic Med) Vol. 7 Issue 5 Pg. e652 (05 2019) ISSN: 2324-9269 [Electronic] United States
• PMID: 30908890 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2019 The Authors. Molecular Genetics & Genomic Medicine published by Wiley Periodicals, Inc.

Chemical References

• GABRB3 protein, human
• Receptors, GABA-A

Topics

• 5' Flanking Region
• Cell Line, Tumor
• HEK293 Cells
• Haplotypes
• Humans
• Promoter Regions, Genetic
• Receptors, GABA-A (genetics, metabolism)
• Schizophrenia (genetics)