A (GATA)7 motif located in the 5' boundary area of the human β-globin locus control region exhibits silencer activity in erythroid cells

Ramani Ramchandran, Chikh Bengra, Barry Whitney, Kenneth Lanclos, Dorothy Tuan Lo

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

A 40-bp DNA, consisting of seven tandem GATA repeats, is located near the HS5 site in the 5' boundary area of the locus control region (LCR) of human β-globin gene. This (GATA)7 motif, named 5a, exhibits silencer activity in erythroid cells. In transfected, recombinant plasmids containing the chloramphenicol acetyltransferase (CAT) reporter gene, 5a repressed the activity of the cis-linked housekeeping phosphoglycerate kinase (pgk) promoter; 5a also repressed the activity of the cis-linked HS2 enhancer regardless of whether the CAT gene was driven by the pgk or the ε-globin promoter. Repression by 5a was most severe when 5a was spliced upstream of HS2 at a distance of less than 200 bases from the HS2 enhancer core. The silencer activity of 5a was independent of whether the component GATA motifs were in head to tail orientation as in the wild type 5a or in head to head or tail to tail orientation as in a mutant 5a. Band shift experiments show that the GATA-1 protein binds to both 5a and the mutant 5a and forms a large protein complex. Together, the results suggest that GATA-1 bound at 5a is a strong, proximal repressor of HS2 enhancer activity. (C) 2000 Wiley-Liss, Inc.

Original languageEnglish (US)
Pages (from-to)14-24
Number of pages11
JournalAmerican Journal of Hematology
Volume65
Issue number1
DOIs
StatePublished - Jan 1 2000

Fingerprint

Locus Control Region
Phosphoglycerate Kinase
Erythroid Cells
Chloramphenicol O-Acetyltransferase
Globins
Tail
Housekeeping
Tandem Repeat Sequences
Reporter Genes
Genes
Proteins
Plasmids
DNA

Keywords

  • GATA-1
  • Globin
  • HS2
  • LCR
  • Silencer

ASJC Scopus subject areas

  • Hematology

Cite this

A (GATA)7 motif located in the 5' boundary area of the human β-globin locus control region exhibits silencer activity in erythroid cells. / Ramchandran, Ramani; Bengra, Chikh; Whitney, Barry; Lanclos, Kenneth; Tuan Lo, Dorothy.

In: American Journal of Hematology, Vol. 65, No. 1, 01.01.2000, p. 14-24.

Research output: Contribution to journalArticle

Ramchandran, Ramani ; Bengra, Chikh ; Whitney, Barry ; Lanclos, Kenneth ; Tuan Lo, Dorothy. / A (GATA)7 motif located in the 5' boundary area of the human β-globin locus control region exhibits silencer activity in erythroid cells. In: American Journal of Hematology. 2000 ; Vol. 65, No. 1. pp. 14-24.
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