Elk-1 protein domains required for direct and SRF-assisted DNA-binding

Nucleic Acids Res. 1992 Jul 11;20(13):3317-24. doi: 10.1093/nar/20.13.3317.

Abstract

The Ets-related Elk-1 protein can bind to purine-rich DNA target sites in a sequence specific fashion and, in addition, can form a ternary complex with the c-fos serum response element (SRE) and the serum response factor (SRF). We demonstrate that Elk-1 can readily interchange between its different interaction partners. The amino terminal ETS-domain of Elk-1 was shown to be necessary and sufficient for direct DNA-binding activity. For ternary complex formation with the SRE and SRF, both the Elk-1 ETS-domain as well as flanking sequences up to amino acid 169 were required. Removal of sequences between the ETS-domain and amino acids 137-169 did not abolish ternary complex formation. This suggests the Elk-1 region spanning amino acids 137-169 to contain a protein-protein interaction domain. Furthermore, we have shown that a single amino acid exchange introduced into the ETS-domain can drastically alter the direct DNA-binding affinity of Elk-1 without severely affecting SRF-assisted binding to the SRE. Thus, Elk-1 requires different propensities of the ETS-domain to exert its different modes of DNA sequence recognition.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites / genetics
  • DNA / metabolism*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism*
  • Genes, fos / genetics
  • HeLa Cells
  • Humans
  • Molecular Sequence Data
  • Nuclear Proteins / genetics
  • Nuclear Proteins / metabolism*
  • Protein-Tyrosine Kinases / chemistry
  • Protein-Tyrosine Kinases / genetics
  • Protein-Tyrosine Kinases / metabolism*
  • Proto-Oncogene Proteins*
  • Retroviridae Proteins, Oncogenic / chemistry
  • Retroviridae Proteins, Oncogenic / genetics
  • Retroviridae Proteins, Oncogenic / metabolism*
  • Serum Response Factor
  • Transcription Factors / genetics
  • Transcription Factors / metabolism*
  • ets-Domain Protein Elk-1

Substances

  • DNA-Binding Proteins
  • ELK1 protein, human
  • Nuclear Proteins
  • Proto-Oncogene Proteins
  • Retroviridae Proteins, Oncogenic
  • Serum Response Factor
  • Transcription Factors
  • ets-Domain Protein Elk-1
  • DNA
  • Protein-Tyrosine Kinases