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Specific RNA binding proteins constructed from zinc fingers

Nature Structural Biology volume 5, pages 543–546 (1998)Cite this article

Abstract

A zinc finger library with degenerate α-helices was displayed on the surface of bacteriophage and proteins that bind human immunodeficiency virus type-1 (HIV-1) Rev response element stem loop IIB (RRE-IIB) RNA or 5S rRNA were isolated. DNA encoding affinity selected zinc fingers was shuffled by recombination in vitro to isolate proteins with higher RNA binding affinity. Proteins constructed in this way bind RNA specifically both in vitro and in vivo . These results demonstrate that RNA substrate specificity of zinc fingers can be changed through mutation of α-helices to construct novel RNA binding proteins.

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Figure 1: Structure and selection of phage displayed zinc fingers.
Figure 2: Compilation of α-helix amino acid sequences of selected zinc fingers.
Figure 3: RNA binding by selected zinc finger peptides in vitro.
Figure 4: Selected zinc finger peptide binding to RRE-IIB RNA in vivo.

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Acknowledgements

We thank E. Rebar and C. Pabo for fd.tet.7000; A. Frankel for pSV2TatRev and pHIV LTR-IIB CAT; T. Parslow for pCH110; and E W. Mercer for cell culture facilities.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Kimmel Cancer Institute, Thomas Jefferson University, 233 S. 10th St., Philadelphia, 19107, Pennsylvania, USA

    Westley J. Friesen & Martyn K. Darby

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  1. Westley J. Friesen
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  2. Martyn K. Darby
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Correspondence to Martyn K. Darby.

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Friesen, W., Darby, M. Specific RNA binding proteins constructed from zinc fingers. Nat Struct Mol Biol 5, 543–546 (1998). https://doi.org/10.1038/794

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