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Titolo:
Binding of IRE-BP to its cognate RNA sequence: SFM studies on a universal RNA backbone for the analysis of RNA-protein interaction
Autore:
Bonin, M; Oberstrass, J; Vogt, U; Wassenegger, M; Nellen, W;
Indirizzi:
Univ Kassel, Genet Abt, D-34132 Kassel, Germany Univ Kassel Kassel Germany D-34132 l, Genet Abt, D-34132 Kassel, Germany Fraunhofer Inst Umweltchem & Okotoxikol, Abt Mol Biotechnol, D-82152 Martinsried, Germany Fraunhofer Inst Umweltchem & Okotoxikol Martinsried Germany D-82152 many
Titolo Testata:
BIOLOGICAL CHEMISTRY
fascicolo: 8, volume: 382, anno: 2001,
pagine: 1157 - 1162
SICI:
1431-6730(200108)382:8<1157:BOITIC>2.0.ZU;2-Y
Fonte:
ISI
Lingua:
ENG
Soggetto:
DOUBLE-STRANDED-RNA; IRON-RESPONSIVE-ELEMENT; ATOMIC-FORCE MICROSCOPY; MONOCLONAL-ANTIBODIES; MESSENGER-RNA; ACONITASE; VISUALIZATION; GENE;
Keywords:
AFM; dsRNA; single molecule analysis;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
19
Recensione:
Indirizzi per estratti:
Indirizzo: Nellen, W Univ Kassel, Genet Abt, Heinrich Plett Str 40, D-34132 Kassel, Germany Univ Kassel Heinrich Plett Str 40 Kassel Germany D-34132 ermany
Citazione:
M. Bonin et al., "Binding of IRE-BP to its cognate RNA sequence: SFM studies on a universal RNA backbone for the analysis of RNA-protein interaction", BIOL CHEM, 382(8), 2001, pp. 1157-1162

Abstract

We have used an RNA consisting of the potato spindle tuber viroid (PSTVd) and 240 bp of double-stranded RNA derived from the GUS gene as a backbone for scanning force microscope (SFM) studies on RNA binding proteins. The in vitro transcribed RNA dforms a rod-like structure of apparent 130 nm in length with a completely base paired central part flanked by the incompletely paired viroid helix with bulges on both sides. The termini of the molecule consist of loops such that no blunt or staggered RNA ends are exposed. Suitable, asymmetrical restriction sites in the construct allow for the insertion of sequences of interest, e. g. protein binding sites. We have inserted the IRE (iron responsive element) sequence into the construct and have usedin vitro transcripts to study binding of IRE-BP. Relative binding frequencies show that 70% of the protein binds to the expected site in the moleculewhile only a slightly enhanced binding is observed at the termini. In the GUS-PSTVd-IRE backbone, the orientation of the molecule is easily determined by IRE-BP binding. It thus provides a versatile tool to study specific aswell as preferential interaction of other proteins with sequences or structures inserted into a different part of the molecule.

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Documento generato il 19/01/20 alle ore 14:38:05