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Cytosine DNA methylation at promoter of non LTR retrotransposon and heat shock protein gene (HSP70) of Entamoeba histolytica and lack of correlation with transcription status
- Source :
- Molecular and Biochemical Parasitology. 212:21-27
- Publication Year :
- 2017
- Publisher :
- Elsevier BV, 2017.
-
Abstract
- Non LTR retrotransposons (EhLINEs and EhSINEs) occupy 11% of the Entamoeba histolytica genome. Since promoter DNA methylation at cytosines has been correlated with transcriptional silencing of transposable elements in model organisms we checked whether this was the case in EhLINE1. We located promoter activity in a 841bp fragment at 5'-end of this element by luciferase reporter assay. From RNAseq and RT-PCR analyses we selected a transcriptionally active and silent copy to study cytosine DNA methylation of the promoter region by bisulfite sequencing. None of the cytosines were methylated in either copy. Further, we looked at methylation status of a few selected cytosines in all 5'-intact EhLINE1 copies by single nucleotide incorporation opposite cytosine in bisulfite-treated DNA, where dGTP would be incorporated if the cytosine was methylated. Again we did not find evidence of cytosine methylation, indicating that expression status of this element was not correlated with promoter DNA methylation. To test for any role of cytosine methylation in transcriptional regulation of the E. histolytica Hsp70 gene in which the promoter is fully methylated under normal growth conditions, we checked methylation status and found that the promoter remained fully methylated during heat-shock as well, although transcription was greatly enhanced by heat-shock, showing that cytosine methylation is not a repressive mark for EhHsp70. Our data present direct evidence that promoter methylation, a common mode of transposon silencing, is unlikely to be involved in transcriptional regulation of EhLINE1, and reinforce the conclusion that promoter DNA methylation may not be a major contributor to transcriptional regulation in E. histolytica.
- Subjects :
- 0301 basic medicine
Retroelements
Transcription, Genetic
Bisulfite sequencing
Biology
Cytosine
03 medical and health sciences
Epigenetics of physical exercise
HSP70 Heat-Shock Proteins
Gene Silencing
Methylated DNA immunoprecipitation
Promoter Regions, Genetic
Molecular Biology
RNA-Directed DNA Methylation
Genetics
030102 biochemistry & molecular biology
Entamoeba histolytica
Promoter
Methylation
DNA Methylation
Molecular biology
Long Interspersed Nucleotide Elements
030104 developmental biology
Gene Expression Regulation
DNA methylation
Illumina Methylation Assay
CpG Islands
Parasitology
Subjects
Details
- ISSN :
- 01666851
- Volume :
- 212
- Database :
- OpenAIRE
- Journal :
- Molecular and Biochemical Parasitology
- Accession number :
- edsair.doi.dedup.....9d1f2c23a6cf552a6d190d2a090811cc