Mutational analysis of the Acropora millepora PaxD paired domain highlights the importance of the linker region for DNA binding

Karin Nordström, Ingo Scholten, Johanna Nordström, Dan Larhammar, David Miller

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Pax transcription factors are found in animals, from simple sponges to insects and vertebrates. The defining feature of Pax proteins is the DNA-binding paired domain (PD), which consists of two helix-turn-helix subdomains, joined with a linker region. Despite high specificity in vivo, the paired domains of different Pax proteins bind similar consensus DNA sequences in vitro. Using bandshift techniques, we show here that the paired domain of the Acropora millepora PaxD protein, which unambiguously belongs to the Pax3/7 group, does not bind to three defined paired domain-binding sites. Domain swapping experiments and site-directed mutagenesis identified two amino acid residues in the linker region of the paired domain as critical to DNA binding; G70 and S71 are highly conserved in Pax proteins, but differ in PaxD (L70 and N71). The PaxD data thus highlight the importance of the linker region, and particularly G70 and S71, in DNA binding by Pax proteins.

Original languageEnglish
Pages (from-to)81-87
Number of pages7
JournalGene
Volume320
Issue number1-2
DOIs
Publication statusPublished - 27 Nov 2003
Externally publishedYes

Keywords

  • Cnidaria
  • EMSA
  • Site-directed mutagenesis

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