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A folding transition and novel zinc finger accessory domain in the transcription factor ADR1

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Abstract

The region responsible for sequence-specific DNA binding by the transcription factor ADR1 contains two Cys2–His2 zinc fingers and an additional N-terminal proximal accessory region (PAR). The N-terminal (non-finger) PAR is unstructured in the absence of DNA and undergoes a folding transition on binding the DNA transcription target site. We have used a set of HN-HN NOEs derived from a perdeuterated protein–DNA complex to describe the fold of ADR1 bound to the UAS1 binding site. The PAR forms a compact domain consisting of three antiparallel strands that contact A-T base pairs in the major groove. The three-strand domain is a novel fold among all known DNA-binding proteins. The PAR shares sequence homology with the N-terminal regions of other zinc finger proteins, suggesting that it represents a new DNA-binding module that extends the binding repertoire of zinc finger proteins.

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Figure 1: The amino acid and nucleotide sequences of the constructs used in these studies.
Figure 2: Constraint distribution and pairwise r.m.s. deviations.
Figure 3: An individual superposition of the calculated family of 25 structures of the first and second zinc fingers of ADR1.
Figure 4: Strip-plots taken from 3D 15N-edited NOESY spectra highlighting residues in the N-terminal region that undergo a folding transition on binding the UAS1.
Figure 5: A backbone alignment of the global fold of the N-terminal region, residues 88–103, as determined by HN-HN NOE contacts.
Figure 6: An NMR spectrum and diagram of the observed and inferred intermolecular contacts.
Figure 7: A model of ADR1-DBD bound to the UAS1, showing both zinc fingers and the N-terminal region.

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Acknowledgements

We thank E.T. Young and C. Rohl for their helpful comments and suggestions in preparing this manuscript. We thank P. Rajagopal for the generous help with NMR experiment implementation and data collection. This work was supported by a grant from the National Institutes of Health. P.M.B. (Department of Biochemistry) was supported by a Public Health Service National Research Award from the National Institute of General Medical Sciences. L.E.S. (Molecular and Cellular Biology Program) was supported by a NIH predoctoral traineeship in Molecular Biophysics.

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  1. Department of Biochemistry, Box 357742, University of Washington, Seattle, 98195-7742 , Washington, USA

    Peter M. Bowers, Lawrence E. Schaufler & Rachel E. Klevit

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  1. Peter M. Bowers
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  3. Rachel E. Klevit
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Correspondence to Rachel E. Klevit.

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Bowers, P., Schaufler, L. & Klevit, R. A folding transition and novel zinc finger accessory domain in the transcription factor ADR1. Nat Struct Mol Biol 6, 478–485 (1999). https://doi.org/10.1038/8283

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