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Sequencing of the rat light neurofilament promoter reveals differences in methylation between expressing and non-expressing cell lines, but not tissues
- Source :
- Gene. 157:325-329
- Publication Year :
- 1995
- Publisher :
- Elsevier BV, 1995.
-
Abstract
- The DNA methylation pattern of the promoter (pNF-L) region of the rat light-neurofilament-encoding gene (NF-L), a neuron-specific gene, was assessed in NF-L expressing and non-expressing cell lines and tissues by genomic sequencing using PCR amplification of bisulfite-modified DNA. We analysed twenty-five potential CpG methylation sites between nucleotide (nt) positions -311 and +103 of pNF-L, containing Sp1- and AP-2-binding sites, a CGCCCCCGC box and a cAMP-responsive element. Six out of 25 possible CpG methylation sites are within these elements. The pNF-L promoter was unmethylated in NF-L-expressing rat brain, as well as in liver not expressing NF-L. In NF-L-expressing PC12 cells, the promoter was unmethylated, whereas in non-expressing glioma C6 cells intensive methylation occurred. A cluster of methylated CpG dinucleotides spanned the region from nt -176 to -67 bp. Thus, methylation of this promoter region could play a role in silencing NF-L in the glioma cell line in vitro, but not in liver tissue in vivo. In a non-CpG sequence, in the CpApG trinucleotide at nt position -114, cytosine was found to be partially methylated. It is thus possible to describe the methylation state of each cytosine present in the area of genomic DNA of interest.
- Subjects :
- Transcription, Genetic
Molecular Sequence Data
Bisulfite sequencing
Gene Expression
Biology
Methylation
Cell Line
03 medical and health sciences
0302 clinical medicine
Epigenetics of physical exercise
Neurofilament Proteins
Sequence Homology, Nucleic Acid
Genetics
Animals
Sulfites
RNA, Messenger
Promoter Regions, Genetic
RNA-Directed DNA Methylation
030304 developmental biology
0303 health sciences
Binding Sites
Base Sequence
DNA
General Medicine
Molecular biology
Rats
Differentially methylated regions
CpG site
DNA methylation
Illumina Methylation Assay
030217 neurology & neurosurgery
Transcription Factors
Subjects
Details
- ISSN :
- 03781119
- Volume :
- 157
- Database :
- OpenAIRE
- Journal :
- Gene
- Accession number :
- edsair.doi.dedup.....81c071fc764d7c6838dc2afc6a8cb6de