Regular Article
DNA Binding and Transactivating Properties of the Paired and Homeobox Protein Pax4

https://doi.org/10.1006/bbrc.1999.0809Get rights and content

Abstract

Transcription factors Pax-4 and Pax-6 are known to be key regulators of pancreatic cell differentiation and development. We report on the cloning of a mouse Pax-4 gene, which contains 10 exons, spanning a 4.7-kbp region. The gene-targeting experiments revealed that Pax-4 and Pax-6 cannot substitute for each other in tissue with overlapping expression of both genes. We identified DNA-binding specificities of Pax-4 paired domain and paired-type homeodomain. Despite the different Pax-4 amino acid residues in positions responsible for Pax-6 paired-domain specificity, the DNA-binding specificities of Pax-4 and Pax-6 are similar. The Pax-4 homeodomain was shown to preferentially dimerize on DNA sequences consisting of an inverted TAAT motif, separated by 4-nucleotide spacing. The Pax-4 transactivation domain was localized within its C-terminal region, which transactivated GAL-based reporter 2.5-fold less than the C-terminal region of Pax-6. We believe that Pax-4 can act as a Pax-6 “repressor,” due to the competition for binding sites and lower transactivation potential of Pax-4.

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    1

    To whom correspondence should be addressed at Institute of Molecular Genetics, Vı́deňská 1083, Prague 4, 142 20, Czech Republic. Fax: (+4202) 44471707. E-mail: [email protected].

    2

    Biochemical Instrumentation Programme, European Molecular Biology Laboratory, Heidelberg, Germany.

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