ArticleRNA (New York, N.Y.)1999
Identification of RNAs that bind to a specific protein using the yeast three-hybrid system.
Article in RNA (New York, N.Y.), 1999. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 69 citations in OpenAlex.
- Decoding RNA-protein interactions using high-throughput methods.RNA biology · 2026Review
- LncRNA SLNCR phenocopies the E2F1 DNA binding site to promote melanoma progression.Cell reports · 2025Article
- Tip110 binding to U6 small nuclear RNA and its participation in pre-mRNA splicing.Cell & bioscience · 2015Article
- Effects of the yeast RNA-binding protein Whi3 on the half-life and abundance of CLN3 mRNA and other targets.PloS one · 2013Article
- Diversity in genetic in vivo methods for protein-protein interaction studies: from the yeast two-hybrid system to the mammalian split-luciferase system.Microbiology and molecular biology reviews : MMBR · 2012Review
- The ribosome assembly factor Nep1 responsible for Bowen-Conradi syndrome is a pseudouridine-N1-specific methyltransferase.Nucleic acids research · 2010Article
- Selections that optimize RNA display in the yeast three-hybrid system.RNA (New York, N.Y.) · 2010Review
- Tandem KH domains of Khd4 recognize AUACCC and are essential for regulation of morphology as well as pathogenicity in Ustilago maydis.RNA (New York, N.Y.) · 2009Article
- Characterization of anti-NF-kappaB RNA aptamer-binding specificity in vitro and in the yeast three-hybrid system.Nucleic acids research · 2009Article
- Whi3, a developmental regulator of budding yeast, binds a large set of mRNAs functionally related to the endoplasmic reticulum.The Journal of biological chemistry · 2008Article
- Combining SELEX and the yeast three-hybrid system for in vivo selection and classification of RNA aptamers.RNA (New York, N.Y.) · 2007Article
- Genetic evidence for 18S rRNA binding and an Rps19p assembly function of yeast nucleolar protein Nep1p.Molecular genetics and genomics : MGG · 2006Article
- A three-hybrid screen identifies mRNAs controlled by a regulatory protein.RNA (New York, N.Y.) · 2006Article
- Musashi regulates the temporal order of mRNA translation during Xenopus oocyte maturation.The EMBO journal · 2006Article
- Recognition of RNA by the p53 tumor suppressor protein in the yeast three-hybrid system.RNA (New York, N.Y.) · 2006Article
- A yeast RNA-hybrid system for the detection of RNA-RNA interactions in vivo.RNA (New York, N.Y.) · 2006Article
- RNA-protein interactions in the yeast three-hybrid system: affinity, sensitivity, and enhanced library screening.RNA (New York, N.Y.) · 2005Article
- RNA sequences that work as transcriptional activating regions.Nucleic acids research · 2003Article
- Having it both ways: transcription factors that bind DNA and RNA.Nucleic acids research · 2002Review
- The RNA-binding specificity of the mouse Dazl protein.Nucleic acids research · 2001Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
Abstract
We have adapted the yeast three-hybrid system to identify RNA ligands for an RNA-binding protein. In this assay system, a protein-RNA interaction is detected by the reconstitution of a transcriptional activator using two hybrid proteins and a hybrid RNA. The RNA molecule is tethered to the promoter of a reporter gene by binding to a hybrid protein consisting of the bacteriophage MS2 coat protein fused to the DNA-binding protein LexA; the RNA-binding domain to be analyzed is fused to the transcriptional activation domain of the yeast Gal4 protein; and the bifunctional RNA consists of binding sites for the coat protein and for the other RNA-binding domain. We built an RNA library such that short fragments of genomic DNA from yeast were transcribed in yeast together with binding sites for the coat protein. We screened this hybrid RNA library for RNAs that bound to the yeast Snp1 protein, a homolog of the human U1-70K protein. The screen yielded as the strongest positive the fragment of U1 RNA that contains loop I, which is known to bind to Snp1 in U1 snRNP. We also identified four other RNA ligands that produced weaker three-hybrid signals, suggesting lower affinities for Snp1 as compared to U1 RNA. In addition, this search also yielded a set of RNA sequences that can activate transcription on their own when bound to a promoter through a protein interaction.
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Registered trials
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