New, Extended Hairpin Form of the Tar-2 Rna Domain Points to the Structural Polymorphism at the 5' End of the Hiv-2 Leader Rna.
From: Laboratory of Structural Chemistry of Nucleic Acids, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznań, Poland.
Nucleic acids research
- Publish Date: 2006
- ISSN: 1362-4962
- Volume: 34
- Issue: 10
- Pages: 2984-97
- Medium: Internet
- Language: English
- Citation (JAMA): Pachulska-Wieczorek Katarzyna, Purzycka Katarzyna J, Adamiak Ryszard W, et al. New, Extended Hairpin Form of the Tar-2 Rna Domain Points to the Structural Polymorphism at the 5' End of the Hiv-2 Leader Rna.. Nucleic Acids Res. 2006;34:2984-97
Abstract
The HIV-2 TAR RNA domain (TAR-2) plays a key role in the trans-activation of HIV-2 transcription as it is the target for the Tat-2 protein and several cell factors. Here, we show that the TAR-2 domain exists in vitro in two global, alternative forms: a new, extended hairpin form with two conformers and the already proposed branched hairpins form. This points strongly to the structural polymorphism of the 5’ end of the HIV-2 leader RNA. The evidence comes from the non-denaturing PAGE mobility assay, 2D structure prediction, enzymatic and Pb2+- or Mg2+-induced RNA cleavages. Existence of the TAR-2 extended form was further proved by the examination of engineered TAR-2 mutants stabilized either in the branched or extended structure. The TAR-2 extended form predominates with an increasing magnesium concentration. Gel retardation assays reveal that both TAR-2 wt and its mutant, unable to form branched structure, bind Tat-2 protein with comparable, high affinity, while RNA hairpins I and II, derived from TAR-2 branched structure model, show much less protein binding. We propose that an internal loop region of the TAR-2 extended hairpin form is a potential Tat-2 binding site.
Mesh Headings (Keywords): 5’ Untranslated Regions, Arginine, Base Sequence, Binding Sites, Diethyl Pyrocarbonate, Electrophoresis, Polyacrylamide Gel, Electrophoretic Mobility Shift Assay, Gene Products, tat, HIV Long Terminal Repeat, HIV-2, Lead, Magnesium, Molecular Sequence Data, Nucleic Acid Conformation, RNA, Viral, Ribonucleases, tat Gene Products, Human Immunodeficiency Virus
Check for Full Text / PubMed Unique Identifier (PMID): 16738137
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