Functional characterization of iron-substituted neural zinc finger factor 1: Metal and DNA binding

Angelique N. Besold, Seung Jae Lee, Sarah L.J. Michel, Niall Lue Sue, Holly J. Cymet

Research output: Contribution to journalArticlepeer-review

Abstract

Neural zinc finger factor 1 (NZF-1) is a nonclassical zinc finger protein involved in neuronal development. NZF-1 contains multiple copies of a unique CCHHC zinc-binding domain that recognize a promoter element in the β-retinoic acid receptor gene termed β-retinoic acid receptor element (β-RARE). Previous studies have established that a two-domain fragment of NZF-1 bound with zinc is sufficient for specific DNA binding. Proper functioning of the nervous system relies heavily on iron and misregulation of this highly redox active metal has serious consequences. Several classes of zinc finger proteins have been shown to bind other metal ions, including iron. To determine if ferrous iron can coordinate to the metal-binding sites of NZF-1 and assess the functional consequences of such coordination, a fragment of NZF-1 that contains two zinc-binding domains, NZF-1 double finger (NZF-1-DF), was prepared. UV-vis spectroscopy experiments demonstrated that Fe(II) is capable of binding to NZF-1-DF. Upon reconstitution with either Fe(II) or Zn(II), NZF-1-DF binds selectively and tightly (nanomolar affinity) to its target β-RARE DNA sequence, whereas apoNZF-1-DF does not bind to DNA and instead aggregates.

Original languageEnglish (US)
Pages (from-to)583-590
Number of pages8
JournalJournal of Biological Inorganic Chemistry
Volume15
Issue number4
DOIs
StatePublished - May 2010
Externally publishedYes

Keywords

  • DNA
  • Fluorescence anisotropy
  • Iron binding
  • Neural zinc finger factor 1
  • Zinc finger

ASJC Scopus subject areas

  • Biochemistry
  • Inorganic Chemistry

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