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- Title
Biophysical characterization of the complex between the iron-responsive transcription factor Fep1 and DNA.
- Authors
Miele, Adriana E.; Cervoni, Laura; Le Roy, Aline; Cutone, Antimo; Musci, Giovanni; Ebel, Christine; Bonaccorsi di Patti, Maria Carmela
- Abstract
Fep1 is an iron-responsive GATA-type transcriptional repressor present in numerous fungi. The DNA-binding domain of this protein is characterized by the presence of two zinc fingers of the Cys2-Cys2 type and a Cys-X5-Cys-X8-Cys-X2-Cys motif located between the two zinc fingers, that is involved in binding of a [2Fe-2S] cluster. In this work, biophysical characterization of the DNA-binding domain of Pichia pastoris Fep1 and of the complex of the protein with cognate DNA has been undertaken. The results obtained by analytical ultracentrifugation sedimentation velocity, small-angle X-ray scattering and differential scanning calorimetry indicate that Fep1 is a natively unstructured protein that is able to bind DNA forming 1:1 and 2:1 complexes more compact than the individual partners. Complex formation takes place independently of the presence of a stoichiometric [2Fe-2S] cluster, suggesting that the cluster may play a role in recruiting other protein(s) required for regulation of transcription in response to changes in intracellular iron levels.
- Subjects
TRANSCRIPTION factors; DNA-binding proteins; SMALL-angle X-ray scattering; PROTEIN domains; DNA; DIFFERENTIAL scanning calorimetry; ZINC-finger proteins
- Publication
European Biophysics Journal, 2021, Vol 50, Issue 3/4, p501
- ISSN
0175-7571
- Publication type
Article
- DOI
10.1007/s00249-020-01489-y