Cargando…

Identification of a novel functional deletion variant in the 5'-UTR of the DJ-1 gene

BACKGROUND: DJ-1 forms part of the neuronal cellular defence mechanism against oxidative insults, due to its ability to undergo self-oxidation. Oxidative stress has been implicated in the pathogenesis of central nervous system damage in different neurodegenerative disorders including Alzheimer'...

Descripción completa

Detalles Bibliográficos
Autores principales: Keyser, Rowena J, van der Merwe, Lize, Venter, Mauritz, Kinnear, Craig, Warnich, Louise, Carr, Jonathan, Bardien, Soraya
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2767350/
https://www.ncbi.nlm.nih.gov/pubmed/19825160
http://dx.doi.org/10.1186/1471-2350-10-105
_version_ 1782173306412072960
author Keyser, Rowena J
van der Merwe, Lize
Venter, Mauritz
Kinnear, Craig
Warnich, Louise
Carr, Jonathan
Bardien, Soraya
author_facet Keyser, Rowena J
van der Merwe, Lize
Venter, Mauritz
Kinnear, Craig
Warnich, Louise
Carr, Jonathan
Bardien, Soraya
author_sort Keyser, Rowena J
collection PubMed
description BACKGROUND: DJ-1 forms part of the neuronal cellular defence mechanism against oxidative insults, due to its ability to undergo self-oxidation. Oxidative stress has been implicated in the pathogenesis of central nervous system damage in different neurodegenerative disorders including Alzheimer's disease and Parkinson's disease (PD). Various mutations in the DJ-1 (PARK7) gene have been shown to cause the autosomal recessive form of PD. In the present study South African PD patients were screened for mutations in DJ-1 and we aimed to investigate the functional significance of a novel 16 bp deletion variant identified in one patient. METHODS: The possible effect of the deletion on promoter activity was investigated using a Dual-Luciferase Reporter assay. The DJ-1 5'-UTR region containing the sequence flanking the 16 bp deletion was cloned into a pGL4.10-Basic luciferase-reporter vector and transfected into HEK293 and BE(2)-M17 neuroblastoma cells. Promoter activity under hydrogen peroxide-induced oxidative stress conditions was also investigated. Computational (in silico) cis-regulatory analysis of DJ-1 promoter sequence was performed using the transcription factor-binding site database, TRANSFAC via the PATCH™ and rVISTA platforms. RESULTS: A novel 16 bp deletion variant (g.-6_+10del) was identified in DJ-1 which spans the transcription start site and is situated 93 bp 3' from a Sp1 site. The deletion caused a reduction in luciferase activity of approximately 47% in HEK293 cells and 60% in BE(2)-M17 cells compared to the wild-type (P < 0.0001), indicating the importance of the 16 bp sequence in transcription regulation. The activity of both constructs was up-regulated during oxidative stress. Bioinformatic analysis revealed putative binding sites for three transcription factors AhR, ARNT, HIF-1 within the 16 bp sequence. The frequency of the g.-6_+10del variant was determined to be 0.7% in South African PD patients (2 heterozygotes in 148 individuals). CONCLUSION: This is the first report of a functional DJ-1 promoter variant, which has the potential to influence transcript stability or translation efficiency. Further work is necessary to determine the extent to which the g.-6_+10del variant affects the normal function of the DJ-1 promoter and whether this variant confers a risk for PD.
format Text
id pubmed-2767350
institution National Center for Biotechnology Information
language English
publishDate 2009
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-27673502009-10-27 Identification of a novel functional deletion variant in the 5'-UTR of the DJ-1 gene Keyser, Rowena J van der Merwe, Lize Venter, Mauritz Kinnear, Craig Warnich, Louise Carr, Jonathan Bardien, Soraya BMC Med Genet Research Article BACKGROUND: DJ-1 forms part of the neuronal cellular defence mechanism against oxidative insults, due to its ability to undergo self-oxidation. Oxidative stress has been implicated in the pathogenesis of central nervous system damage in different neurodegenerative disorders including Alzheimer's disease and Parkinson's disease (PD). Various mutations in the DJ-1 (PARK7) gene have been shown to cause the autosomal recessive form of PD. In the present study South African PD patients were screened for mutations in DJ-1 and we aimed to investigate the functional significance of a novel 16 bp deletion variant identified in one patient. METHODS: The possible effect of the deletion on promoter activity was investigated using a Dual-Luciferase Reporter assay. The DJ-1 5'-UTR region containing the sequence flanking the 16 bp deletion was cloned into a pGL4.10-Basic luciferase-reporter vector and transfected into HEK293 and BE(2)-M17 neuroblastoma cells. Promoter activity under hydrogen peroxide-induced oxidative stress conditions was also investigated. Computational (in silico) cis-regulatory analysis of DJ-1 promoter sequence was performed using the transcription factor-binding site database, TRANSFAC via the PATCH™ and rVISTA platforms. RESULTS: A novel 16 bp deletion variant (g.-6_+10del) was identified in DJ-1 which spans the transcription start site and is situated 93 bp 3' from a Sp1 site. The deletion caused a reduction in luciferase activity of approximately 47% in HEK293 cells and 60% in BE(2)-M17 cells compared to the wild-type (P < 0.0001), indicating the importance of the 16 bp sequence in transcription regulation. The activity of both constructs was up-regulated during oxidative stress. Bioinformatic analysis revealed putative binding sites for three transcription factors AhR, ARNT, HIF-1 within the 16 bp sequence. The frequency of the g.-6_+10del variant was determined to be 0.7% in South African PD patients (2 heterozygotes in 148 individuals). CONCLUSION: This is the first report of a functional DJ-1 promoter variant, which has the potential to influence transcript stability or translation efficiency. Further work is necessary to determine the extent to which the g.-6_+10del variant affects the normal function of the DJ-1 promoter and whether this variant confers a risk for PD. BioMed Central 2009-10-13 /pmc/articles/PMC2767350/ /pubmed/19825160 http://dx.doi.org/10.1186/1471-2350-10-105 Text en Copyright © 2009 Keyser et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Keyser, Rowena J
van der Merwe, Lize
Venter, Mauritz
Kinnear, Craig
Warnich, Louise
Carr, Jonathan
Bardien, Soraya
Identification of a novel functional deletion variant in the 5'-UTR of the DJ-1 gene
title Identification of a novel functional deletion variant in the 5'-UTR of the DJ-1 gene
title_full Identification of a novel functional deletion variant in the 5'-UTR of the DJ-1 gene
title_fullStr Identification of a novel functional deletion variant in the 5'-UTR of the DJ-1 gene
title_full_unstemmed Identification of a novel functional deletion variant in the 5'-UTR of the DJ-1 gene
title_short Identification of a novel functional deletion variant in the 5'-UTR of the DJ-1 gene
title_sort identification of a novel functional deletion variant in the 5'-utr of the dj-1 gene
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2767350/
https://www.ncbi.nlm.nih.gov/pubmed/19825160
http://dx.doi.org/10.1186/1471-2350-10-105
work_keys_str_mv AT keyserrowenaj identificationofanovelfunctionaldeletionvariantinthe5utrofthedj1gene
AT vandermerwelize identificationofanovelfunctionaldeletionvariantinthe5utrofthedj1gene
AT ventermauritz identificationofanovelfunctionaldeletionvariantinthe5utrofthedj1gene
AT kinnearcraig identificationofanovelfunctionaldeletionvariantinthe5utrofthedj1gene
AT warnichlouise identificationofanovelfunctionaldeletionvariantinthe5utrofthedj1gene
AT carrjonathan identificationofanovelfunctionaldeletionvariantinthe5utrofthedj1gene
AT bardiensoraya identificationofanovelfunctionaldeletionvariantinthe5utrofthedj1gene