NMR and ICP spectroscopic analysis of the DNA-binding domain of the Drosophila GCM protein reveals a novel Zn2+-binding motif

Masato Shimizu, Hidekazu Hiroaki, Daisuke Kohda, Toshihiko Hosoya, Yasuko Akiyama-Oda, Yoshiki Hotta, Eugene Hayato Morita, Kosuke Morikawa

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    2 Citations (Scopus)

    Abstract

    Drosophila GCM (glial cell missing) is a novel DNA-binding protein that determines the fate of glial precursors from the neural default to glia. The GCM protein contains the functional domain that is essential for recognition of the upstream sequence of the repo gene. In the DNA-binding region of this GCM protein, there is a cysteine-rich region with which divalent metal ions such as Zn2+ must bind and other proteins belonging to the GCM family have a corresponding region. To obtain a more detailed insight into the structural and functional features of this DNA-binding region, we have determined the minimal DNA-binding domain and obtained inductively coupled plasma atomic emission spectra and 1H-15N, 1H-15N-13C and 113Cd2+ NMR spectra, with or without its specific DNA molecule. Considering the results, it was concluded that the minimal DNA-binding domain includes two Zn2+-binding sites, one of which is adjacent to the interface for DNA binding. Systematic mutational analyses of the conserved cysteine residues in the minimal DNA-binding domain revealed that one Zn2+-binding site is indispensable for stabilization of the higher order structure of this DNA-binding domain, but that the other is not.

    Original languageEnglish
    Pages (from-to)247-254
    Number of pages8
    JournalProtein Engineering
    Volume16
    Issue number4
    Publication statusPublished - Apr 1 2003

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    All Science Journal Classification (ASJC) codes

    • Biochemistry
    • Molecular Biology

    Cite this

    Shimizu, M., Hiroaki, H., Kohda, D., Hosoya, T., Akiyama-Oda, Y., Hotta, Y., ... Morikawa, K. (2003). NMR and ICP spectroscopic analysis of the DNA-binding domain of the Drosophila GCM protein reveals a novel Zn2+-binding motif. Protein Engineering, 16(4), 247-254.