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PDBsum entry 2ab7
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RNA binding protein
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PDB id
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2ab7
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References listed in PDB file
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Key reference
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Title
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Solution structures and characterization of human immunodeficiency virus rev responsive element iib RNA targeting zinc finger proteins.
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Authors
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S.H.Mishra,
C.M.Shelley,
D.J.Barrow,
M.K.Darby,
M.W.Germann.
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Ref.
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Biopolymers, 2006,
83,
352-364.
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PubMed id
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Abstract
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The Rev responsive element (RRE), a part of unspliced human immunodeficiency
virus (HIV) RNA, serves a crucial role in the production of infectious HIV
virions. The viral protein Rev binds to RRE and facilitates transport of mRNA to
the cytoplasm. Inhibition of the Rev-RRE interaction disrupts the viral life
cycle. Using a phage display protocol, dual zinc finger proteins (ZNFs) were
generated that bind specifically to RREIIB at the high affinity Rev binding
site. These proteins were further shortened and simplified, and they still
retained their RNA binding affinity. The solution structures of ZNF29 and a
mutant, ZNF29G29R, have been determined by nuclear magnetic resonance (NMR)
spectroscopy. Both proteins form C(2)H(2)-type zinc fingers with essentially
identical structures. RNA protein interactions were evaluated quantitatively by
isothermal titration calorimetry, which revealed dissociation constants (K(d)'s)
in the nanomolar range. The interaction with the RNA is dependent upon the zinc
finger structure; in the presence of EDTA, RNA binding is abolished. For both
proteins, RNA binding is mediated by the alpha-helical portion of the zinc
fingers and target the bulge region of RREIIB-TR. However, ZNF29G29R exhibits
significantly stronger binding to the RNA target than ZNF29; this illustrates
that the binding of the zinc finger scaffold is amenable to further improvements.
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