Binding of A/T-rich DNA by three high mobility group-like domains in c-Abl tyrosine kinase

J Biol Chem. 1996 Sep 13;271(37):22823-30. doi: 10.1074/jbc.271.37.22823.

Abstract

The c-Abl tyrosine kinase has been shown previously to bind DNA. Using polymerase chain reaction-based binding site-selection methods, no consensus high affinity binding site for c-Abl was found. Instead, oligonucleotides with runs of A/T sequences were isolated, and purified c-Abl was shown to bind A/T-containing oligonucleotides better than those without A/T sequences. DNA binding of c-Abl was dependent on three high mobility group 1-like boxes (HLBs), which bound cooperatively to the A/T-rich oligonucleotides. To distinguish binding to A/T sequences per se from binding to nonspecific DNA with a bend at the A/T-rich region, two oligonucleotides were compared for binding to c-Abl. Both oligonucleotides contained A/T sequences. In one, the A/T motif was part of an 80-mer duplex DNA. In another, the A/T motif was in the duplex arm of an 80-mer "bubble DNA" containing an internal unpaired 20-mer region to provide a flexible hinge. Interestingly, the HLBs of c-Abl bound better to the oligonucleotide containing the bubble, suggesting a higher affinity for bent DNA rather than A/T sequences per se. Taken together, these observations define a new class of DNA binding domains, the HLBs, which do not bind DNA with a high degree of sequence specificity, but may selectively bind to bent DNA or to sequences that are easier to distort.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine / metabolism
  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Binding Sites
  • Cell Line
  • DNA / metabolism*
  • High Mobility Group Proteins / metabolism*
  • Mice
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Proto-Oncogene Proteins c-abl / metabolism*
  • Sequence Alignment
  • Thymidine / metabolism

Substances

  • High Mobility Group Proteins
  • DNA
  • Proto-Oncogene Proteins c-abl
  • Adenosine
  • Thymidine