Nuclear protein-binding sites in a transcriptional control region of the rabbit α-globin gene

S. E. Yost, B. Shewchuk, R. Hardison

Research output: Contribution to journalArticlepeer-review

19 Scopus citations

Abstract

The 5'-flanking and internal regions of the rabbit α-globin gene, which constitute a CpG island, are required for enhancer-independent expression in transfected cells. In this study, electrophoretic mobility shift assays revealed that a battery of nuclear proteins from both erythroid and nonerythroid cells bind specifically to these regulatory regions. Assays based on exonuclease III digestion, methylation interference, and DNase I footprinting identified sequences bound by proteins in crude nuclear extracts and by purified transcription factor Sp1. In the 5' flank, recognition sites for the transcription factors α-IRP (positions -53 to -44 relative to the cap site), CP1 (-73 to -69), and Sp1 (-95 to -90) are bound by proteins in K562 cell nuclear extracts, as are three extended upstream regions. Two recognition sites for Sp1 in intron 1 are also bound both by proteins in crude nuclear extracts and by purified Sp1. The sequences CCAC in intron 2 and C5 in the 3'-untranslated region also bind proteins. A major binding site found in exon 1, TATGGCGC, matches in sequence and methylation interference pattern the binding site for nuclear protein YY1, and binding is inhibited through competition by YY1-specific oligonucleotides. The protein- binding sites flanking and internal to the rabbit α-globin gene may form an extended promoter.

Original languageEnglish (US)
Pages (from-to)5439-5449
Number of pages11
JournalMolecular and cellular biology
Volume13
Issue number9
DOIs
StatePublished - 1993

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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