Specific RNA binding proteins constructed from zinc fingers.
A zinc finger library with degenerate alpha-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 alpha-helices to construct novel RNA binding proteins.
Department of Microbiology and Immunology, Kimmel Cancer Institute, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
W J
WJ
Friesen
M K
MK
Darby
D000595:Amino Acid Sequence; D001483:Base Sequence; D015694:Gene Products, rev / Q000378:metabolism; D015497:HIV-1; D006801:Humans; D008969:Molecular Sequence Data; D019151:Peptide Library; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012341:RNA, Ribosomal, 5S / Q000737:chemistry*; D012367:RNA, Viral / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000737:chemistry*; D016335:Zinc Fingers; D054321:rev Gene Products, Human Immunodeficiency Virus
D015694:Gene Products, rev; D019151:Peptide Library; D012341:RNA, Ribosomal, 5S; D012367:RNA, Viral; D016601:RNA-Binding Proteins; D054321:rev Gene Products, Human Immunodeficiency Virus