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Binding specificity of human nuclear protein interacting with the Alu-family DNA repeats

Biochem Biophys Res Commun. 1991 Jul 31;178(2):613-9. doi: 10.1016/0006-291x(91)90152-w.

Abstract

We have demonstrated earlier that human cells contain nuclear protein interacting with conserved GC-rich sequence motifs of human Alu-family DNA repeats. One of these sequences is located in the region between elements A and B of bipartite RNA polymerase III promoter of Alu (AB-region). In this study we have used a DNase I footprinting assay with an Alu restriction subfragment covering AB-region, as well as a gel mobility shift assay with appropriate synthetic oligonucleotides to analyse in more detail the interaction of the protein with AB-region. We have also used antibodies raised against a zinc-finger peptide to examine the presence of a zinc-finger in the Alu-binding protein. The results indicate that AGG triplets may be important for high-affinity binding of the protein to DNA, and that the Alu-binding protein is a zinc-finger protein.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Binding, Competitive
  • DNA / genetics
  • DNA / metabolism*
  • Humans
  • Molecular Sequence Data
  • Nuclear Proteins / metabolism*
  • Oligonucleotide Probes
  • Promoter Regions, Genetic*
  • RNA Polymerase III / genetics*
  • Repetitive Sequences, Nucleic Acid
  • Restriction Mapping
  • Zinc Fingers / genetics

Substances

  • Nuclear Proteins
  • Oligonucleotide Probes
  • DNA
  • RNA Polymerase III