2ab7
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(New page: 200px<br /> <applet load="2ab7" size="450" color="white" frame="true" align="right" spinBox="true" caption="2ab7" /> '''Solution structures and characterization of...)
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Revision as of 12:34, 8 November 2007
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Solution structures and characterization of HIV RRE IIB RNA targeting zinc finger proteins
Overview
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.
About this Structure
2AB7 is a Protein complex structure of sequences from Escherichia coli with ZN as ligand. Full crystallographic information is available from OCA.
Reference
Solution structures and characterization of human immunodeficiency virus Rev responsive element IIB RNA targeting zinc finger proteins., Mishra SH, Shelley CM, Barrow DJ Jr, Darby MK, Germann MW, Biopolymers. 2006 Nov;83(4):352-64. PMID:16826557
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