DNA affinity cleaving analysis of homeodomain-DNA interaction: Identification of homeodomain consensus sites in genomic DNA

Zhigang Shang, Yon W. Ebright, Nancy Iler, P. Shannon Pendergrast, Yann Echelard, Andrew P. Mcmahon, Richard H. Ebright, Cory Abate

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

We have incorporated the DNA-cleaving moiety o-phenanthroline-copper at amino acid 10 of the Msx-1 homeodomain, and we have analyzed site-specific DNA cleavage by the resulting Msx-1 derivative. We show that amino acid 10 of the Msx-1 homeodomain is close to the 5' end of the consensus DNA site 5'- (C/G)TAATTG-3' in the Msx-1-DNA complex. Our results indicate that the orientation of the Msx-1 homeodomain relative to DNA is analogous to the orientation of the engrailed and Antennapedia homeodomains. We show further that DNA affinity cleaving permits identification of consensus DNA sites for Msx-1 in kilobase DNA substrates. The specificity of the approach enabled us to identify an Msx-1 consensus DNA site within the transcriptional control region of the developmental regulatory gene Wnt-1. We propose that incorporation of o-phenanthroline-copper at amino acid 10 of a homeodomain may provide a generalizable strategy to determine the orientation of a homeodomain relative to DNA and to identify homeodomain consensus DNA sites in genomic DNA.

Original languageEnglish (US)
Pages (from-to)118-122
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume91
Issue number1
DOIs
StatePublished - Jan 4 1994

All Science Journal Classification (ASJC) codes

  • General

Keywords

  • Msx-1
  • Wnt-1
  • cysteine-specific chemical modification of protein
  • o-phenanthroline-copper

Fingerprint

Dive into the research topics of 'DNA affinity cleaving analysis of homeodomain-DNA interaction: Identification of homeodomain consensus sites in genomic DNA'. Together they form a unique fingerprint.

Cite this