Protein binding elements in the human β-polymerase promoter

Ella Englander, Samuel H. Wilson

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The core promoter for human DNA polymerase β contains discrete binding sites for mammalian nuclear proteins, as revealed by DNasel footprinting and gel mobility shift assays. Two sites correspond to sequences identical with the Sp1 factor binding element, and a third site includes an eight residue palindromic sequence, TGACGTCA, known as the CRE element of several cAMP responsive promoters; the 5 to 10 residues flanking this palindrome on each side have no apparent sequence homology with known elements in other promoters. Nuclear extract from a variety of tissues and cells were examined; these included rat liver and testes and cultured cells of human and hamster origin. The DNasel footprint is strong over and around the palindromic element for each of the extracts and is equivalent in size (∼22 residues); footprinting over the Sp1 binding sites is seen also. Two potential tissue-specific binding sites, present in liver but not in testes, were found corresponding to residues -13 to -10 and +33 to +48, respectively. Protein binding to the palindromic element was confirmed by an electrophoretic mobility shift assay with the core promoter as probe. Binding specificity of the 22 residue palindromic element, as revealed by oligonucleotide competition, is different from that of AP-1 binding element. Controlled proteolysis with trypsin was used to study structural properties of proteins forming the mobility shift bands. Following digestion with trypsin, most of the palindrome binding activity of each extract corresponded to a sharp, faster migrating band, potentially representing a DNA binding domain of the palindrome binding protein.

Original languageEnglish (US)
Pages (from-to)919-928
Number of pages10
JournalNucleic Acids Research
Volume18
Issue number4
StatePublished - Feb 25 1990
Externally publishedYes

Fingerprint

Binding sites
Promoter
Protein Binding
Binding Sites
Electrophoretic Mobility Shift Assay
Protein
Liver
Trypsin
Testis
Assays
DNA
Tissue
Proteolysis
Electrophoretic mobility
Palindrome
Oligonucleotides
Transcription Factor AP-1
DNA-Directed DNA Polymerase
Sequence Homology
Nuclear Proteins

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Protein binding elements in the human β-polymerase promoter. / Englander, Ella; Wilson, Samuel H.

In: Nucleic Acids Research, Vol. 18, No. 4, 25.02.1990, p. 919-928.

Research output: Contribution to journalArticle

Englander, Ella ; Wilson, Samuel H. / Protein binding elements in the human β-polymerase promoter. In: Nucleic Acids Research. 1990 ; Vol. 18, No. 4. pp. 919-928.
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