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PMID: 8355692 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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

Molecular and cellular biology ·Vol. 13 ·No. 9 ·1993-09-00 ·Pages 5439-49

Yost SE, Shewchuk B, Hardison R

Abstract

The 5'-flanking and internal regions of the rabbit alpha-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 alpha-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 alpha-globin gene may form an extended promoter.

MeSH Terms
Animals Base Sequence Binding Sites Gene Expression Regulation Genes Globins/genetics Goats Humans In Vitro Techniques Mice Molecular Sequence Data Nuclear Proteins/metabolism Oligodeoxyribonucleotides/chemistry Promoter Regions, Genetic Rabbits Sequence Alignment Sp1 Transcription Factor/metabolism Tumor Cells, Cultured
Chemicals
Nuclear Proteins Oligodeoxyribonucleotides Sp1 Transcription Factor Globins
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Yost S E
Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.
Shewchuk B
Hardison R
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.13.9.5439-5449.1993
Published
1993-09-00
Pages
5439-49
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC360253
Subset
IM
Grants
NIDDK NIH HHS · DK-01589 · United States
NIDDK NIH HHS · DK-27635 · United States
NHLBI NIH HHS · HL-44491 · United States
External Links
PubMed source DOI: 10.1128/mcb.13.9.5439-5449.1993 PMC Full Text View journal details
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