Methylation of α-type embryonic globin gene απ represses transcription in primary erythroid cells

Rakesh Singal, Jane M. VanWert, Larry Ferdinand

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The inverse relationship between expression and methylation of β-type globin genes is well established. However, little is known about the relationship between expression and methylation of avian α-type globin genes. The embryonic απ-globin promoter was unmethylated, and α-globin RNA was easily detected in 5-day chicken erythroid cells. A progressive methylation of the CpG dinucleotides in the απ promoter associated with loss of expression of απ-globin gene was seen during development in primary erythroid cells. A 315-bp απ-globin promoter region was cloned in an expression construct (απpGL3E) containing a luciferase reporter gene and SV40 enhancer. The απpGL3E construct was transfected into primary erythroid cells derived from 5-day-old chicken embryos. Methylation of απpGL3E plasmid and απ-globin promoter alone resulted in a 20-fold and 7-fold inhibition of expression, respectively. The fully methylated but not the unmethylated 315-bp απ-globin gene promoter fragment formed a methyl cytosinebinding protein complex (MeCPC). Chromatin immunoprecipitation assays were combined with quantitative real-time polymerase chain reaction to assess histone acetylation associated with the απ-globin gene promoter. Slight hyperacetylation of histone H3 but a marked hyperacetylation of histone H4 was seen in 5-day when compared with 14-day erythroid cells. These results demonstrate that methylation can silence transcription of an avian α-type embryonic globin gene in homologous primary erythroid cells, possibly by interacting with an MeCPC and histone deacetylase complex.

Original languageEnglish
Pages (from-to)4217-4222
Number of pages6
JournalBlood
Volume100
Issue number12
DOIs
StatePublished - Dec 1 2002
Externally publishedYes

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Erythroid Cells
Methylation
Globins
Transcription
Genes
Histones
Chickens
Acetylation
Histone Deacetylases
Chromatin Immunoprecipitation
Polymerase chain reaction
Luciferases
Reporter Genes
Genetic Promoter Regions
Chromatin
Real-Time Polymerase Chain Reaction
Assays
Proteins
Plasmids
Embryonic Structures

ASJC Scopus subject areas

  • Hematology

Cite this

Methylation of α-type embryonic globin gene απ represses transcription in primary erythroid cells. / Singal, Rakesh; VanWert, Jane M.; Ferdinand, Larry.

In: Blood, Vol. 100, No. 12, 01.12.2002, p. 4217-4222.

Research output: Contribution to journalArticle

Singal, Rakesh ; VanWert, Jane M. ; Ferdinand, Larry. / Methylation of α-type embryonic globin gene απ represses transcription in primary erythroid cells. In: Blood. 2002 ; Vol. 100, No. 12. pp. 4217-4222.
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