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Effect of allelic variation at the NACP-Rep1 repeat upstream of the alpha-synuclein gene (SNCA) on transcription in a cell culture luciferase reporter system.

Publication ,  Journal Article
Chiba-Falek, O; Nussbaum, RL
Published in: Hum Mol Genet
December 15, 2001

Mutations in the alpha-synuclein gene (SNCA) have been implicated in familial Parkinson's disease (PD) while certain polymorphic alleles at a microsatellite repeat, NACP-Rep1, located approximately 10 kb upstream of the gene, have been associated with sporadic PD. In order to study the regulation of the human alpha-synuclein gene, we performed a deletion analysis of 10.7 kb upstream of the translational start site, using the luciferase reporter assay in 293T cells and the neuroblastoma cell line SH-SY5Y. The shortest fragment, 400 bp upstream of the transcriptional start site, was sufficient for transcription in both cell lines. The other constructs led to variable expression levels, with some showing maximum expression and others showing nearly complete extinction of expression. An 880 bp fragment located approximately 10 kb upstream of the gene and containing the NACP-Rep1 polymorphism, was shown to be necessary for normal expression. Additional analysis of the NACP-Rep1 locus and surrounding DNA suggested that two domains flanking the repeat interact to enhance expression while the repeat acts as a negative modulator. Next, we measured the activity of the entire 10.7 kb upstream region in the luciferase reporter assay when each of our different NACP-Rep1 alleles were present. The expression levels varied very significantly among the different alleles over a 3-fold range in the SH-SY5Y cells but showed little or no significant variation in the 293T cells. Given that even small changes in alpha-synuclein expression may, over many decades, predispose to PD, the association of different NACP-Rep1 alleles with PD may be a consequence of polymorphic differences in transcriptional regulation of alpha-synuclein expression resulting from different NACP-Rep1 alleles.

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Published In

Hum Mol Genet

DOI

ISSN

0964-6906

Publication Date

December 15, 2001

Volume

10

Issue

26

Start / End Page

3101 / 3109

Location

England

Related Subject Headings

  • alpha-Synuclein
  • Transcription, Genetic
  • Synucleins
  • Repetitive Sequences, Nucleic Acid
  • Promoter Regions, Genetic
  • Polymorphism, Genetic
  • Phosphoproteins
  • Parkinson Disease
  • Nerve Tissue Proteins
  • Luciferases
 

Citation

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Chiba-Falek, O., & Nussbaum, R. L. (2001). Effect of allelic variation at the NACP-Rep1 repeat upstream of the alpha-synuclein gene (SNCA) on transcription in a cell culture luciferase reporter system. Hum Mol Genet, 10(26), 3101–3109. https://doi.org/10.1093/hmg/10.26.3101
Chiba-Falek, O., and R. L. Nussbaum. “Effect of allelic variation at the NACP-Rep1 repeat upstream of the alpha-synuclein gene (SNCA) on transcription in a cell culture luciferase reporter system.Hum Mol Genet 10, no. 26 (December 15, 2001): 3101–9. https://doi.org/10.1093/hmg/10.26.3101.
Chiba-Falek, O., and R. L. Nussbaum. “Effect of allelic variation at the NACP-Rep1 repeat upstream of the alpha-synuclein gene (SNCA) on transcription in a cell culture luciferase reporter system.Hum Mol Genet, vol. 10, no. 26, Dec. 2001, pp. 3101–09. Pubmed, doi:10.1093/hmg/10.26.3101.
Journal cover image

Published In

Hum Mol Genet

DOI

ISSN

0964-6906

Publication Date

December 15, 2001

Volume

10

Issue

26

Start / End Page

3101 / 3109

Location

England

Related Subject Headings

  • alpha-Synuclein
  • Transcription, Genetic
  • Synucleins
  • Repetitive Sequences, Nucleic Acid
  • Promoter Regions, Genetic
  • Polymorphism, Genetic
  • Phosphoproteins
  • Parkinson Disease
  • Nerve Tissue Proteins
  • Luciferases