Structural and Dynamics Characterization of the MerR Family Metalloregulator CueR in its Repression and Activation States

Hila Sameach, Aya Narunsky, Salome Azoulay-Ginsburg, Lada Gevorkyan-Aiapetov, Yonathan Zehavi, Yoni Moskovitz, Tamar Juven-Gershon, Nir Ben-Tal, Sharon Ruthstein

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

CueR (Cu export regulator) is a metalloregulator protein that “senses” Cu(I) ions with very high affinity, thereby stimulating DNA binding and the transcription activation of two other metalloregulator proteins. The crystal structures of CueR when unbound or bound to DNA and a metal ion are very similar to each other, and the role of CueR and Cu(I) in initiating the transcription has not been fully understood yet. Using double electron-electron resonance (DEER) measurements and structure modeling, we investigate the conformational changes that CueR undergoes upon binding Cu(I) and DNA in solution. We observe three distinct conformations, corresponding to apo-CueR, DNA-bound CueR in the absence of Cu(I) (the “repression” state), and CueR-Cu(I)-DNA (the “activation” state). We propose a detailed structural mechanism underlying CueR's regulation of the transcription process. The mechanism explicitly shows the dependence of CueR activity on copper, thereby revealing the important negative feedback mechanism essential for regulating the intracellular copper concentration.

Original languageEnglish
Pages (from-to)988-996.e3
JournalStructure
Volume25
Issue number7
DOIs
StatePublished - 5 Jul 2017

Bibliographical note

Publisher Copyright:
© 2017 Elsevier Ltd

Funding

S.R. acknowledges the support of MC CIG, grant no. 303636 and ISF grant no. 176/16. N.B.-T. acknowledges the support of grant no. 1775/12 of the I-CORE Program of the Planning and Budgeting Committee and The Israel Science Foundation. A.N. was funded in part by the Edmond J. Safra Center for Bioinformatics at Tel Aviv University. The Elexsys E580 Bruker EPR spectrometer was partially supported by the Israel Science Foundation, grant no. 564/12.

FundersFunder number
Edmond J. Safra Center for Bioinformatics
MC CIG303636
Israel Science Foundation1775/12, 176/16
Tel Aviv University564/12
Israeli Centers for Research Excellence

    Keywords

    • ConTemplate
    • CueR
    • DEER
    • EPR
    • MerR family metalloregulators
    • protein dynamics
    • protein-DNA interaction
    • site-directed spin labeling

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