Elsevier

Gene

Volume 808, 15 January 2022, 145973
Gene

Research paper
The effect of human GRIN1 gene 5′ functional region on gene expression regulation in vitro

https://doi.org/10.1016/j.gene.2021.145973Get rights and content
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open access

Highlights

  • −337 to −159 bp and −704 to −556 bp of GRIN1 contain transcription repression regions.

  • −556 to −337 bp of GRIN1 enhanced gene expression in neuronal cell lines.

  • The proximal promoter region of GRIN1 may related to neuropsychiatric diseases.

Abstract

Introduction

Abnormal expression of ionotropic glutamate receptor NMDA type subunit 1, the key subunit of the NMDA receptor, may be related to many neuropsychiatric disorders. In this study, we explored the functional sequence of the 5′ regulatory region of the human GRIN1 gene and discussed the transcription factors that may regulate gene expression.

Materials and Methods

Twelve recombinant pGL3 vectors with gradually truncated fragment lengths were constructed, transfected into HEK-293, U87, and SK-N-SH cell lines, and analyzed through the luciferase reporter gene assay. JASPAR database is used to predict transcription factors.

Results

In SK-N-SH and U87 cell lines, regions from −337 to −159 bp, −704 to −556 bp inhibited gene expression, while −556 to −337 bp upregulated gene expression. In HEK-293 and U87 cell lines, the expression of fragment −1703 to + 188 bp was significantly increased compared to adjacent fragments −1539 to + 188 bp and −1843 to + 188 bp. The protein expressions of fragments −2162 to + 188 bp and −2025 to + 188 bp, −1539 to + 188 bp and −1215 to + 188 bp, −1215 to + 188 bp and −1066 to + 188 bp were significantly different in HEK-293 and SK-N-SH cells. According to the predictions of the JASPAR database, the transcription factors REST, EGR1, and CREB1/HIC2 may bind the DNA sequences of GRIN1 gene from the −337 to −159, −556 to −337, and −704 to −556, respectively. In addition, zinc finger transcription factors may regulate the expression of other differentially expressed fragments.

Conclusions

Abnormal transcription regulation in the proximal promoter region of GRIN1 (-704 to + 188 bp) may be involved in the course of neuropsychiatric diseases.

Abbreviations

NMDA receptors
ionotropic glutamate N-methyl-D-aspartate receptors
NRSE
neuron-restrictive silencing element
P19 cells
undifferentiated embryonic terato-carcinoma cells
PC12 cells
phaeochromocytoma cells
HEK-293 cells
human embryonic kidney cell
U87 cells
human glioblastoma
SK-N-SH cells
human neuroblastoma cell
LUC
firefly luciferase
TK
renilla luciferase
NRSF
neuron-restrictive silencing factor
REST
RE1-silencing transcription factor
EGR1
early growth response protein-1
PFC
prefrontal cortex
CREB1
cAMP response element-binding protein
bZIP
basic leucine zipper family
HIC2
hypermethylated in cancer 2

Keywords

GRIN1
Transcriptional regulation
Promoter
Neuropsychiatric diseases

Cited by (0)

1

Equal contribution.