PUBLICATION
The Upstream 1350~1250 Nucleotide Sequences of the Human ENDOU-1 Gene Contain Critical Cis-Elements Responsible for Upregulating Its Transcription during ER Stress
- Authors
- Lee, H.C., Chao, H.T., Lee, S.Y., Lin, C.Y., Tsai, H.J.
- ID
- ZDB-PUB-231224-26
- Date
- 2023
- Source
- International Journal of Molecular Sciences 24(24): (Journal)
- Registered Authors
- Lee, Hung-Chieh, Lin, Cheng-Yung, Tsai, Huai-Jen
- Keywords
- ENDOU-1 gene, ER stress, HEK-293T, cis-acting element, promoter analysis, transcription factor, transcriptional regulation, zebrafish
- MeSH Terms
-
- Animals
- Base Sequence
- Gene Expression Regulation
- Humans
- Promoter Regions, Genetic
- Transcription, Genetic
- Uridylate-Specific Endoribonucleases*/genetics
- Zebrafish*/genetics
- PubMed
- 38139221 Full text @ Int. J. Mol. Sci.
Citation
Lee, H.C., Chao, H.T., Lee, S.Y., Lin, C.Y., Tsai, H.J. (2023) The Upstream 1350~1250 Nucleotide Sequences of the Human ENDOU-1 Gene Contain Critical Cis-Elements Responsible for Upregulating Its Transcription during ER Stress. International Journal of Molecular Sciences. 24(24):.
Abstract
ENDOU-1 encodes an endoribonuclease that overcomes the inhibitory upstream open reading frame (uORF)-trap at 5'-untranslated region (UTR) of the CHOP transcript, allowing the downstream coding sequence of CHOP be translated during endoplasmic reticulum (ER) stress. However, transcriptional control of ENDOU-1 remains enigmatic. To address this, we cloned an upstream 2.1 kb (-2055~+77 bp) of human ENDOU-1 (pE2.1p) fused with reporter luciferase (luc) cDNA. The promoter strength driven by pE2.1p was significantly upregulated in both pE2.1p-transfected cells and pE2.1p-injected zebrafish embryos treated with stress inducers. Comparing the luc activities driven by pE2.1p and -1125~+77 (pE1.2p) segments, we revealed that cis-elements located at the -2055~-1125 segment might play a critical role in ENDOU-1 upregulation during ER stress. Since bioinformatics analysis predicted many cis-elements clustered at the -1850~-1250, we further deconstructed this segment to generate pE2.1p-based derivatives lacking -1850~-1750, -1749~-1650, -1649~-1486, -1485~-1350 or -1350~-1250 segments. Quantification of promoter activities driven by these five internal deletion plasmids suggested a repressor binding element within the -1649~-1486 and an activator binding element within the -1350~-1250. Since luc activities driven by the -1649~-1486 were not significantly different between normal and stress conditions, we herein propose that the stress-inducible activator bound at the -1350~-1250 segment makes a major contribution to the increased expression of human ENDOU-1 upon ER stresses.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping