Developmentally regulated demethylase activity targeting the βA-globin gene in primary avian erythroid cells

Kavitha Ramachandran, Jane Van Wert, Gopal Gopisetty, Rakesh Singal

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Differential expression of globin genes has provided an interesting model system for better understanding commonly inherited diseases such as thalassemia. In the avian β-type globin cluster (5′-ρ-βH- βA-ε-3′), silencing of the embryonic ρ-globin gene occurs concomitantly with the activation of the adult βA-globin gene during embryonic development. DNA methylation is a dynamic process that regulates gene expression. We observed a progressive loss of methylation of βA-globin gene, during avian embryonic development that was concurrent with the expression of the gene. The promoter and exon 1 regions of the template strand were completely demethylated, whereas residual methylation was retained in exons 2 and 3. Using a modified methylation-sensitive single-nucleotide primer extension (MS-SNuPE) assay, we observed stage-specific demethylase activity in the nuclear extracts of chicken red cells; activity in 5-, 8-, and 11-day-old erythroid cell nuclear extracts was 6, 76, and 24%, respectively. The demethylase targeted both hemimethylated and fully methylated substrates. Our findings demonstrate stage-specific demethylase activity in nuclear extracts from primary chicken erythroid cells that could target the fully methylated promoter of a developmentally regulated native gene.

Original languageEnglish
Pages (from-to)3416-3422
Number of pages7
JournalBiochemistry
Volume46
Issue number11
DOIs
StatePublished - Mar 20 2007

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Erythroid Cells
Globins
Genes
Methylation
Gene Expression
Embryonic Development
Chickens
Exons
Thalassemia
DNA Methylation
Cell Extracts
Gene expression
Nucleotides
Assays
Chemical activation
Cells
Substrates

ASJC Scopus subject areas

  • Biochemistry

Cite this

Developmentally regulated demethylase activity targeting the βA-globin gene in primary avian erythroid cells. / Ramachandran, Kavitha; Van Wert, Jane; Gopisetty, Gopal; Singal, Rakesh.

In: Biochemistry, Vol. 46, No. 11, 20.03.2007, p. 3416-3422.

Research output: Contribution to journalArticle

Ramachandran, Kavitha ; Van Wert, Jane ; Gopisetty, Gopal ; Singal, Rakesh. / Developmentally regulated demethylase activity targeting the βA-globin gene in primary avian erythroid cells. In: Biochemistry. 2007 ; Vol. 46, No. 11. pp. 3416-3422.
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