Structural defects of a Pax8 mutant that give rise to congenital hypothyroidism

Gianluca Tell, Lucia Pellizzari, Gennaro Esposito, Carlo Pucillo, Paolo Emidio Macchia, Roberto Di Lauro, Giuseppe Damante

    Research output: Contribution to journalArticle

    Abstract

    Pax proteins are transcriptional regulators that play important roles during embryogenesis. These proteins recognize specific DNA sequences via a conserved element: the paired domain (Prd domain). The low level of organized secondary structure, in the free state, is a general feature of Prd domains; however, these proteins undergo a dramatic gain in α-helical content upon interaction with DNA ('induced fit'). Pax8 is expressed in the developing thyroid, kidney and several areas of the central nervous system. In humans, mutations of the Pax8 gene, which are mapped to the coding region of the Prd domain, give rise to congenital hypothyroidism. Here, we have investigated the molecular defects caused by a mutation in which leucine at position 62 is substituted for an arginine. Leu62 is conserved among Prd domains, and contributes towards the packing together of helices 1 and 3. The binding amnity of the Leu62Arg mutant for a specific DNA sequence (the C sequence of thyroglobulin promoter) is decreased 60-fold with respect to the wildtype Pax8 Prd domain. However, the affinities with which the wild-type and the mutant proteins bind to a non-specific DNA sequence are very similar. CD spectra demonstrate that, in the absence ofDNA, both wild-type Pax8 and the Leu62Arg mutant possess a low α-helical content; however, in the Leu62Arg mutant, the gain in α-helical content upon interaction with DNA is greatly reduced with respect to the wild-type protein. Thus the molecular defect of the Leu62Arg mutant causes a reduced capability for induced fit upon DNA interaction.

    Original languageEnglish (US)
    Pages (from-to)89-93
    Number of pages5
    JournalBiochemical Journal
    Volume341
    Issue number1
    DOIs
    StatePublished - Jul 1 1999

    Fingerprint

    Congenital Hypothyroidism
    DNA sequences
    Defects
    DNA
    Proteins
    Mutation
    Thyroglobulin
    Neurology
    Mutant Proteins
    Leucine
    Embryonic Development
    Arginine
    Thyroid Gland
    Central Nervous System
    Genes
    Kidney

    Keywords

    • DNA binding
    • Human mutation
    • Induced fit
    • Pax genes
    • Protein structure

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    Tell, G., Pellizzari, L., Esposito, G., Pucillo, C., Macchia, P. E., Di Lauro, R., & Damante, G. (1999). Structural defects of a Pax8 mutant that give rise to congenital hypothyroidism. Biochemical Journal, 341(1), 89-93. https://doi.org/10.1042/0264-6021:3410089

    Structural defects of a Pax8 mutant that give rise to congenital hypothyroidism. / Tell, Gianluca; Pellizzari, Lucia; Esposito, Gennaro; Pucillo, Carlo; Macchia, Paolo Emidio; Di Lauro, Roberto; Damante, Giuseppe.

    In: Biochemical Journal, Vol. 341, No. 1, 01.07.1999, p. 89-93.

    Research output: Contribution to journalArticle

    Tell, G, Pellizzari, L, Esposito, G, Pucillo, C, Macchia, PE, Di Lauro, R & Damante, G 1999, 'Structural defects of a Pax8 mutant that give rise to congenital hypothyroidism', Biochemical Journal, vol. 341, no. 1, pp. 89-93. https://doi.org/10.1042/0264-6021:3410089
    Tell G, Pellizzari L, Esposito G, Pucillo C, Macchia PE, Di Lauro R et al. Structural defects of a Pax8 mutant that give rise to congenital hypothyroidism. Biochemical Journal. 1999 Jul 1;341(1):89-93. https://doi.org/10.1042/0264-6021:3410089
    Tell, Gianluca ; Pellizzari, Lucia ; Esposito, Gennaro ; Pucillo, Carlo ; Macchia, Paolo Emidio ; Di Lauro, Roberto ; Damante, Giuseppe. / Structural defects of a Pax8 mutant that give rise to congenital hypothyroidism. In: Biochemical Journal. 1999 ; Vol. 341, No. 1. pp. 89-93.
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