2AB3

Solution structures and characterization of HIV RRE IIB RNA targeting zinc finger proteins

2AB3 の概要

• エントリーDOI: 10.2210/pdb2ab3/pdb
• 分子名称: ZNF29, ZINC ION (2 entities in total)
• 機能のキーワード: zinc finger protein, beta beta alpha, rreiib-tr, rna binding protein
• 由来する生物種: Escherichia coli
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 3622.56

構造登録者

Mishra, S.H.,Shelley, C.M.,Darby, M.K.,Germann, M.W. (登録日: 2005-07-14, 公開日: 2005-08-02, 最終更新日: 2024-05-22)

主引用文献

Mishra, S.H.,Shelley, C.M.,Barrow, D.J.,Darby, M.K.,Germann, M.W.
Solution structures and characterization of human immunodeficiency virus Rev responsive element IIB RNA targeting zinc finger proteins.
Biopolymers, 83:352-364, 2006

PubMed Abstract: 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.

PubMed: 16826557
DOI: 10.1002/bip.20565

実験手法

SOLUTION NMR