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 journalArticlepeer-review


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
Issue number1
StatePublished - Jul 1 1999


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

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


Dive into the research topics of 'Structural defects of a Pax8 mutant that give rise to congenital hypothyroidism'. Together they form a unique fingerprint.

Cite this