First Author | Iler N | Year | 1996 |
Journal | Biochem Biophys Res Commun | Volume | 227 |
Issue | 1 | Pages | 257-65 |
PubMed ID | 8858134 | Mgi Jnum | J:35778 |
Mgi Id | MGI:83223 | Doi | 10.1006/bbrc.1996.1498 |
Citation | Iler N, et al. (1996) Rapid identification of homeodomain binding sites in the Wnt-5a gene using an immunoprecipitation strategy. Biochem Biophys Res Commun 227(1):257-65 |
abstractText | Here we describe an immunoprecipitation approach for identifying homeodomain binding sites within uncharacterized genomic sequences of a putative downstream target gene, Wnt-5a. Immunoprecipitation of Wnt-5a genomic fragments was performed using a purified Msx1 homeodomain polypeptide (Msx1) and its corresponding antisera (alpha-Msx1). This resulted in isolation of three fragments containing multiple DNA binding sites for Msx1, as confirmed by DNA binding studies. The three fragments were contiguous within a 3.4 kb intronic sequence of Wnt-5a. Moreover, at least one of the Msx1 sites has been conserved throughout evolution, suggesting that these sites may comprise or contribute to a regulatory element for Wnt-5a. We propose that the immunoprecipitation strategy permits a rapid, initial approach for identifying functionally-relevant homeodomain binding sites within target genes whose regulatory sequences have not yet been previously elucidated. |