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Association between perinatal methylation of the neuronal differentiation regulator HES1 and later childhood neurocognitive function and behaviour.

Lillycrop KA, Costello PM, Teh AL, Murray RJ, Clarke-Harris R, Barton SJ, Garratt ES, Ngo S, Sheppard AM, Wong J, Dogra S, Burdge GC, Cooper C, Inskip HM, Gale CR, Gluckman PD, Harvey NC, Chong YS, Yap F, Meaney MJ, Rifkin-Graboi A, Holbrook JD, Epigen Global Research ConsortiumGodfrey KM - Int J Epidemiol (2015)

Bottom Line: Consistent with these findings, higher HES1 methylation was associated with higher executive memory function in a second independent group of 200 SWS 7-year-olds.Here, HES1 DMROI methylation predicted differences in early infant behaviour, known to be associated with academic success.Thus, our findings suggest that perinatal epigenetic processes mark later neurocognitive function and behaviour, providing support for a role of epigenetic processes in mediating the long-term consequences of early life environment on cognitive development.

View Article: PubMed Central - PubMed

Affiliation: Centre for Biological Sciences, and NIHR Southampton Biomedical Research Centre, University Hospital Southampton, Southampton, UK, kal@soton.ac.uk.

No MeSH data available.


Related in: MedlinePlus

Methylation of CpG5 blocks ETS transcription factor binding to the HES1 promoter sequence. Results are typical of three analyses. (a) The unmethylated biotin-labelled probe showed a strong shift upon incubation with nuclear extract from the human neuroblastoma cell line IMR32; this shift was markedly reduced by co-incubating with 500-fold excess of the unlabelled specific competitor, but not with 500-fold excess of an unlabelled non-specific competitor. (b) Binding to the probe was markedly diminished by co-incubation with 100-fold excess of an unlabelled oligonucleotide containing the core consensus sequence for ETS (GGAA) but not with 100-fold excess of a mutated ETS core competitor (c) The unmethylated probe was incubated with 50-, 100- and 500-fold excesses of the unmethylated or methylated competitor; binding to the unmethylated probe was competed out with a 100-fold excess of the methylated competitor compared with a 500-fold excess of the unmethylated competitor.
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dyv052-F4: Methylation of CpG5 blocks ETS transcription factor binding to the HES1 promoter sequence. Results are typical of three analyses. (a) The unmethylated biotin-labelled probe showed a strong shift upon incubation with nuclear extract from the human neuroblastoma cell line IMR32; this shift was markedly reduced by co-incubating with 500-fold excess of the unlabelled specific competitor, but not with 500-fold excess of an unlabelled non-specific competitor. (b) Binding to the probe was markedly diminished by co-incubation with 100-fold excess of an unlabelled oligonucleotide containing the core consensus sequence for ETS (GGAA) but not with 100-fold excess of a mutated ETS core competitor (c) The unmethylated probe was incubated with 50-, 100- and 500-fold excesses of the unmethylated or methylated competitor; binding to the unmethylated probe was competed out with a 100-fold excess of the methylated competitor compared with a 500-fold excess of the unmethylated competitor.

Mentions: To determine whether methylation of these CpG loci had functional consequences by influencing transcription factor binding to the HES1 promoter, electrophoretic mobility shift assays (EMSAs) were used. In the human neuroblastoma cell line IMR32, one specific protein complex bound to the HES1 promoter −4706 to −4740 region, containing CpG sites 2–5 (Figure 4a). In silico analysis of this region of the HES1 promoter using the Predict transcription factor binding sites (PROMO) software31 predicted that CpG5 was located within an ELK1 (part of the ETS family) binding site. Multiplexed consensus competitor EMSAs32 identified the transcription factor bound at this site as part of the ETS transcription factor family. This was confirmed by specific competitive binding with an ETS consensus sequence but not an oligonucleotide containing a mutated core ‘GGAA’ ETS binding sequence (Figure 4b). Moreover, wheras binding was substantially reduced in the presence of 100-fold excess of unmethylated specific competitor, it was unaffected in the presence of 100-fold excess of a specific competitor sequence containing methylated CpG5 (Figure 4c). This suggests that ETS binds preferentially to the unmethylated sequence upstream of HES1 and methylation of CpG5 inhibits ETS binding to this locus.Figure 4.


Association between perinatal methylation of the neuronal differentiation regulator HES1 and later childhood neurocognitive function and behaviour.

Lillycrop KA, Costello PM, Teh AL, Murray RJ, Clarke-Harris R, Barton SJ, Garratt ES, Ngo S, Sheppard AM, Wong J, Dogra S, Burdge GC, Cooper C, Inskip HM, Gale CR, Gluckman PD, Harvey NC, Chong YS, Yap F, Meaney MJ, Rifkin-Graboi A, Holbrook JD, Epigen Global Research ConsortiumGodfrey KM - Int J Epidemiol (2015)

Methylation of CpG5 blocks ETS transcription factor binding to the HES1 promoter sequence. Results are typical of three analyses. (a) The unmethylated biotin-labelled probe showed a strong shift upon incubation with nuclear extract from the human neuroblastoma cell line IMR32; this shift was markedly reduced by co-incubating with 500-fold excess of the unlabelled specific competitor, but not with 500-fold excess of an unlabelled non-specific competitor. (b) Binding to the probe was markedly diminished by co-incubation with 100-fold excess of an unlabelled oligonucleotide containing the core consensus sequence for ETS (GGAA) but not with 100-fold excess of a mutated ETS core competitor (c) The unmethylated probe was incubated with 50-, 100- and 500-fold excesses of the unmethylated or methylated competitor; binding to the unmethylated probe was competed out with a 100-fold excess of the methylated competitor compared with a 500-fold excess of the unmethylated competitor.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4588869&req=5

dyv052-F4: Methylation of CpG5 blocks ETS transcription factor binding to the HES1 promoter sequence. Results are typical of three analyses. (a) The unmethylated biotin-labelled probe showed a strong shift upon incubation with nuclear extract from the human neuroblastoma cell line IMR32; this shift was markedly reduced by co-incubating with 500-fold excess of the unlabelled specific competitor, but not with 500-fold excess of an unlabelled non-specific competitor. (b) Binding to the probe was markedly diminished by co-incubation with 100-fold excess of an unlabelled oligonucleotide containing the core consensus sequence for ETS (GGAA) but not with 100-fold excess of a mutated ETS core competitor (c) The unmethylated probe was incubated with 50-, 100- and 500-fold excesses of the unmethylated or methylated competitor; binding to the unmethylated probe was competed out with a 100-fold excess of the methylated competitor compared with a 500-fold excess of the unmethylated competitor.
Mentions: To determine whether methylation of these CpG loci had functional consequences by influencing transcription factor binding to the HES1 promoter, electrophoretic mobility shift assays (EMSAs) were used. In the human neuroblastoma cell line IMR32, one specific protein complex bound to the HES1 promoter −4706 to −4740 region, containing CpG sites 2–5 (Figure 4a). In silico analysis of this region of the HES1 promoter using the Predict transcription factor binding sites (PROMO) software31 predicted that CpG5 was located within an ELK1 (part of the ETS family) binding site. Multiplexed consensus competitor EMSAs32 identified the transcription factor bound at this site as part of the ETS transcription factor family. This was confirmed by specific competitive binding with an ETS consensus sequence but not an oligonucleotide containing a mutated core ‘GGAA’ ETS binding sequence (Figure 4b). Moreover, wheras binding was substantially reduced in the presence of 100-fold excess of unmethylated specific competitor, it was unaffected in the presence of 100-fold excess of a specific competitor sequence containing methylated CpG5 (Figure 4c). This suggests that ETS binds preferentially to the unmethylated sequence upstream of HES1 and methylation of CpG5 inhibits ETS binding to this locus.Figure 4.

Bottom Line: Consistent with these findings, higher HES1 methylation was associated with higher executive memory function in a second independent group of 200 SWS 7-year-olds.Here, HES1 DMROI methylation predicted differences in early infant behaviour, known to be associated with academic success.Thus, our findings suggest that perinatal epigenetic processes mark later neurocognitive function and behaviour, providing support for a role of epigenetic processes in mediating the long-term consequences of early life environment on cognitive development.

View Article: PubMed Central - PubMed

Affiliation: Centre for Biological Sciences, and NIHR Southampton Biomedical Research Centre, University Hospital Southampton, Southampton, UK, kal@soton.ac.uk.

No MeSH data available.


Related in: MedlinePlus