The state of DNA methylation in the promoter regions of the human red cell membrane protein (band 3, protein 4.2, andβ-spectrin) genes

Abstract

Indrikis Muiznieks 1,5The state of methylation of the 5Ј-CpG-3Ј sites is known to be linked to the regulation of Birgit Schmitz 1 promoter function by modulating DNA-protein interactions and to the structure of chromaGudrun Schell 1 tin. As part of a project to determine methylation patterns in the human genome, the Yoshihito Yawata 2 methylation profiles were examined in genes for the human erythroid membrane proteins; Walter Doerfler 1 protein 4.2 (P4.2), gene (ELB42), band 3 (B3), gene (EPB3), and β-spectrin (β-Sp), gene 1 Institute of Genetics, University (SPTB). The bisulfite protocol of the genomic sequencing method was applied.(1) In the of Cologne, Köln, Germany DNA from peripheral white blood cells, the promoter regions of EPB3 and ELB42 were 2 Division of Hematology, extensively methylated, but the promoter of SPTB was totally unmethylated. (2) During Department of Medicine, erythroid differentiation, (i) ELB42 was unmethylated in DNAs from the cell line UT-7/EPO, Kawasaki Medical School, but became methylated (55Ϫ93%) in cultured erythroblasts from peripheral BFU-E. The Kurashiki City, Japan mRNA from ELB42 was first detected in early erythroblasts and protein 4.2 was expressed Present addresses: in late erythroblasts. (ii) EPB3 was consistently methylated in UT-7/EPO and also in cultu-3 Institute of Human Genetics red erythroblasts from burst forming unit erythroid (BFU-E) from peripheral blood. EPB3 and Anthropology, Heinrichand ELB42 were efficiently transcribed in UT-7 cells only after erythropoietin stimulation. Heine-University, Duesseldorf, (iii) SPTB remained unmethylated in DNAs from UT-7/EPO and cultured erythroblasts. (3) Germany 4 Institute of Human Genetics, We also investigated methylation profiles in peripheral white blood cells from patients with University of Essen Medical erythroid diseases, like complete P4.2 deficiency due to ELB42 mutations, hereditary School, Essen, Germany spherocytosis with EPB3 mutations, and hereditary elliptocytosis with SPTB mutations. 5 Department of Microbiology,The methylation profiles of the promoter regions of these three genes were essentially Latvian University, Riga, Latvia identical to those in healthy individuals.

Genotypic and phenotypic expressions of protein 4.2 in human erythroid cells

“…The number of the 5'-CpG-3' dinucleotide sites appears to be small, unlike the β-spectrin gene [14], which has numerous 5'-CpG-3' sites as so-called ''CpG islands'' [15−20]. The 5'-CpG-3' sites of the protein 4.2 gene were highly methylated in mononuclear cells from normal human peripheral blood [21].…”

Genotypic and phenotypic expressions of protein 4.2 in human erythroid cells

“…The number of the 5'-CpG-3' dinucleotide sites appears to be small, unlike the β-spectrin gene [14], which has numerous 5'-CpG-3' sites as so-called ''CpG islands'' [15−20]. The 5'-CpG-3' sites of the protein 4.2 gene were highly methylated in mononuclear cells from normal human peripheral blood [21].…”

On the Biological Significance of DNA Methylation

“…These structural alterations of the erythrocyte membrane are responsible for hematologic diseases like hereditary elliptocytosis or hereditary spherocytosis [106]. By the bisulfite genomic sequencing procedure we determined patterns of methylation in the promoter and 5′ regions of the following human genes: the protein 4.2 (P4.2) gene (ELB42), the band 3 (B3) gene (EPB3), and the β spectrin (β Sp) gene (SPTB) [25,26]. The promot er regions of the EPB3 and ELB42 genes were extensively methylated, whereas the promoters of the SPTB and the ankyrin genes were unmethylated.…”

“…5 2005 with the adenovirus system could reliably guide all our efforts. In collaboration with several groups, we deter mined patterns of DNA methylation in the Prader-Willi/Angelman regions of the human genome [20,21], in the promoter regions of the RET protoonco gene [22], of the FMR1 gene [23,24], and of several genes of the erythrocyte membrane [25,26].…”

Foreign DNA Integration—Perturbations of the Genome—Oncogenesis

“…This more general role was thought to be in the establishment of chromatin domains for which patterns of DNA methylation might present the initial scaffold on which to build more complex structures by specific DNA‐protein interactions. A number of human genomic segments were therefore investigated for specific DNA methylation patterns 49–58. Patterns of methylation were highly specific for different parts of the human genome and were different from cell type to cell type.…”

“…Patterns of methylation were highly specific for different parts of the human genome and were different from cell type to cell type. The patterns of DNA methylation in the human genome were frequently found to be highly conserved between individuals 49–57…”

Genotypic and phenotypic expressions of protein 4.2 in human erythroid cells

“…The number of the 5'-CpG-3' dinucleotide sites appears to be small, unlike the β-spectrin gene [14], which has numerous 5'-CpG-3' sites as so-called ''CpG islands'' [15−20]. The 5'-CpG-3' sites of the protein 4.2 gene were highly methylated in mononuclear cells from normal human peripheral blood [21].…”

On the Biological Significance of DNA Methylation

“…These structural alterations of the erythrocyte membrane are responsible for hematologic diseases like hereditary elliptocytosis or hereditary spherocytosis [106]. By the bisulfite genomic sequencing procedure we determined patterns of methylation in the promoter and 5′ regions of the following human genes: the protein 4.2 (P4.2) gene (ELB42), the band 3 (B3) gene (EPB3), and the β spectrin (β Sp) gene (SPTB) [25,26]. The promot er regions of the EPB3 and ELB42 genes were extensively methylated, whereas the promoters of the SPTB and the ankyrin genes were unmethylated.…”

“…5 2005 with the adenovirus system could reliably guide all our efforts. In collaboration with several groups, we deter mined patterns of DNA methylation in the Prader-Willi/Angelman regions of the human genome [20,21], in the promoter regions of the RET protoonco gene [22], of the FMR1 gene [23,24], and of several genes of the erythrocyte membrane [25,26].…”

Foreign DNA Integration—Perturbations of the Genome—Oncogenesis

“…This more general role was thought to be in the establishment of chromatin domains for which patterns of DNA methylation might present the initial scaffold on which to build more complex structures by specific DNA‐protein interactions. A number of human genomic segments were therefore investigated for specific DNA methylation patterns 49–58. Patterns of methylation were highly specific for different parts of the human genome and were different from cell type to cell type.…”

“…Patterns of methylation were highly specific for different parts of the human genome and were different from cell type to cell type. The patterns of DNA methylation in the human genome were frequently found to be highly conserved between individuals 49–57…”

Genotypic and phenotypic expressions of protein 4.2 in human erythroid cells

“…The number of the 5'-CpG-3' dinucleotide sites appears to be small, unlike the β-spectrin gene [14], which has numerous 5'-CpG-3' sites as so-called ''CpG islands'' [15−20]. The 5'-CpG-3' sites of the protein 4.2 gene were highly methylated in mononuclear cells from normal human peripheral blood [21].…”

On the Biological Significance of DNA Methylation

“…These structural alterations of the erythrocyte membrane are responsible for hematologic diseases like hereditary elliptocytosis or hereditary spherocytosis [106]. By the bisulfite genomic sequencing procedure we determined patterns of methylation in the promoter and 5′ regions of the following human genes: the protein 4.2 (P4.2) gene (ELB42), the band 3 (B3) gene (EPB3), and the β spectrin (β Sp) gene (SPTB) [25,26]. The promot er regions of the EPB3 and ELB42 genes were extensively methylated, whereas the promoters of the SPTB and the ankyrin genes were unmethylated.…”

“…5 2005 with the adenovirus system could reliably guide all our efforts. In collaboration with several groups, we deter mined patterns of DNA methylation in the Prader-Willi/Angelman regions of the human genome [20,21], in the promoter regions of the RET protoonco gene [22], of the FMR1 gene [23,24], and of several genes of the erythrocyte membrane [25,26].…”

Foreign DNA Integration—Perturbations of the Genome—Oncogenesis

“…This more general role was thought to be in the establishment of chromatin domains for which patterns of DNA methylation might present the initial scaffold on which to build more complex structures by specific DNA‐protein interactions. A number of human genomic segments were therefore investigated for specific DNA methylation patterns 49–58. Patterns of methylation were highly specific for different parts of the human genome and were different from cell type to cell type.…”

“…Patterns of methylation were highly specific for different parts of the human genome and were different from cell type to cell type. The patterns of DNA methylation in the human genome were frequently found to be highly conserved between individuals 49–57…”