Dissecting protein:protein interactions between transcription factors with an RNA aptamer.

Important note: Information in this article was accurate in 1996. The state of the art may have changed since the publication date.

Dissecting protein:protein interactions between transcription factors with an RNA aptamer.

RNA. 1995 May;1(3):317-26. Unique Identifier : AIDSLINE MED/96079963
Tian Y; Adya N; Wagner S; Giam CZ; Green MR; Ellington AD; Department of Chemistry, Indiana University, Bloomington 47405,; USA.

Abstract: Nucleic acid aptamers isolated from random sequence pools have generally proven useful at inhibiting the interactions of nucleic acid binding proteins with their cognate nucleic acids. In order to develop reagents that could also be used to study protein:protein interactions, we have used in vitro selection to search for RNA aptamers that could interact with the transactivating protein Tax from human T-cell leukemia virus. Tax does not normally bind to nucleic acids, but instead stimulates transcription by interacting with a variety of cellular transcription factors, including the cyclic AMP-response element binding protein (CREB), NF-kappa B, and the serum response factor (SRF). Starting from a pool of greater than 10(13) different RNAs with a core of 120 random sequence positions, RNAs were selected for their ability to be co-retained on nitrocellulose filters with Tax. After five cycles of selection and amplification, a single nucleic acid species remained. This aptamer was found to bind Tax with high affinity and specificity, and could disrupt complex formation between Tax and NF-kappa B, but not with SRF. The differential effects of our aptamer probe on protein:protein interactions suggest a model for how the transcription factor binding sites on the surface of the Tax protein are organized. This model is consistent with data from a variety of other studies.

Keywords: Base Sequence Binding Sites Binding, Competitive DNA-Binding Protein, Cyclic AMP-Responsive/METABOLISM DNA-Binding Proteins/METABOLISM Gene Products, tax/ANTAGONISTS & INHIB/*METABOLISM Molecular Sequence Data Nuclear Proteins/METABOLISM NF-kappa B/METABOLISM Protein Binding RNA/ANTAGONISTS & INHIB/*METABOLISM Support, Non-U.S. Gov't Support, U.S. Gov't, Non-P.H.S. Transcription Factors/*METABOLISM JOURNAL ARTICLE
960330
M9630726

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