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Volumn 9, Issue 1, 1998, Pages 90-96

Macromolecular matchmaking: Advances in two-hybrid and related technologies

Author keywords

[No Author keywords available]

Indexed keywords

DNA; LIGAND; PROTEIN; RNA;

EID: 0031908472     PISSN: 09581669     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0958-1669(98)80090-6     Document Type: Article
Times cited : (34)

References (44)
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    • of special interest. Inouye et al. have come up with a modest but useful modification of the original two-hybrid system that facilitates the study of the interaction of a protein simultaneously with two partners. The system employs two separate bait-binding domain fusions which bind to distinct DNA elements engineered into independent reporter genes. Interaction of an activation domain fusion with each of the binding domain fusions can be followed by monitoring the differential activities of the two associated reporters.
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    • of special interest. A reverse two-hybrid system is described that can be used to detect mutations or molecules that disrupt a protein - protein or protein - DNA interaction of interest. The URA3 counterselectable marker replaces the HIS3 reporter, such that a two-hybrid interaction results in sensitivity to 5-fluoroorotic acid. Dissociation of the interaction leads to 5-fluoroorotic acid resistance, providing a positive selection scheme for the loss of an interaction. See Huang and Schreiber 1998 [41] for a description of a similar, somewhat improved system.
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    • of special interest. Similar to the work presented by Vidal et al. 1996 [6], Shin et al. have developed a different positive genetic selection scheme for mutations, proteins, peptides, or drugs that disrupt protein - protein interactions. See Leanna and Hannick 1996 [40] for a description of a third such system.
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    • of special interest. P.L. Bartel, Fields S. 1 Oxford University Press New York This first edition of The Yeast Two-Hybrid System provides a fairly comprehensive and detailed overview of the two-hybrid system and related technologies. Chapter 13 discusses the specifics of a two-hybrid system which is performed in mammalian cells
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    • Accelerated aging and nucleolar fragmentation in yeast sgs1 mutants
    • of special interest. Many genes in yeast and mammals encode very similar proteins. The value of yeast as a model system is best shown by human disease genes of which little is known beyond the fact that their inheritance results in disease. Werner's syndrome is a disease with several hallmarks of premature aging and a cellular phenotype characterized by a reduced life-span in culture. The sequence of the human gene was found to be highly similar to that of the yeast SGS1 gene, which encodes a DNA helicase. In this remarkable report, Sinclair et al. show that SGS1 mutant yeast cell have a markedly reduced life-span and share other cellular phenotypes with cells from individuals with Werner's syndrome. This paper clearly demonstrates the utility of using yeast cells as a living test-tube to facilitate the study of mechanisms of human disease.
    • Sinclair DA, Mills K, Guarente L. Accelerated aging and nucleolar fragmentation in yeast sgs1 mutants. of special interest Science. 277:1997;1313-1316 Many genes in yeast and mammals encode very similar proteins. The value of yeast as a model system is best shown by human disease genes of which little is known beyond the fact that their inheritance results in disease. Werner's syndrome is a disease with several hallmarks of premature aging and a cellular phenotype characterized by a reduced life-span in culture. The sequence of the human gene was found to be highly similar to that of the yeast SGS1 gene, which encodes a DNA helicase. In this remarkable report, Sinclair et al. show that SGS1 mutant yeast cell have a markedly reduced life-span and share other cellular phenotypes with cells from individuals with Werner's syndrome. This paper clearly demonstrates the utility of using yeast cells as a living test-tube to facilitate the study of mechanisms of human disease.
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    • A three-hybrid system for detecting small ligand - Protein receptor interactions
    • of outstanding interest. Licitra and Liu describe the use of a heterodimeric ligand (dexamethasone - FK506) to link together two-hybrid DNA-binding and activation domains fused to the corresponding ligand-binding domains. They then used this three-hybrid system to isolate overlapping clones of the FK506 binding protein FKBP12, from a cDNA-activation domain fusion library when screened in the presence of the ligand. This system should prove useful to identify receptors for novel ligands and to screen synthetic compounds produced by combinatorial chemistry for interaction with a target protein.
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    • Inducible gene expression and protein translocation using nontoxic ligands identified by a mammalian three-hybrid screen
    • of special interest of outstanding interest. Rapamycin has proven to be a useful reagent to control protein dimerization in vivo, due to its ability to interact with two proteins simultaneously (FRAP and FKBP12). An important limitation on its use has been its inhibitory effect on cell growth, mediated by FRAP. To get around this problem, these workers have synthesized a chemically altered rapamycin, and using three-hybrid technology described by Belshaw et al. 1996 [20] and Licitra and Liu 1996 [21], have identified mutations in the ligand-binding domain of FRAP that permit binding to the modified rapamycin. The work described in this paper is an elegant example of the potential utility of three-hybrid technology
    • Liberles SD, Diver ST, Austin DJ, Schreiber SL. Inducible gene expression and protein translocation using nontoxic ligands identified by a mammalian three-hybrid screen. of special interest of outstanding interest Proc Natl Acad Sci USA. 94:1997;7825-7830 Rapamycin has proven to be a useful reagent to control protein dimerization in vivo, due to its ability to interact with two proteins simultaneously (FRAP and FKBP12). An important limitation on its use has been its inhibitory effect on cell growth, mediated by FRAP. To get around this problem, these workers have synthesized a chemically altered rapamycin, and using three-hybrid technology described by Belshaw et al. 1996 [20] and Licitra and Liu 1996 [21], have identified mutations in the ligand-binding domain of FRAP that permit binding to the modified rapamycin. The work described in this paper is an elegant example of the potential utility of three-hybrid technology.
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    • Liberles, S.D.1    Diver, S.T.2    Austin, D.J.3    Schreiber, S.L.4
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    • of special interest. Amara et al. describe the development of next generation synthetic dimerizers, based on ligands for FKBP-12. The ultimate aim is to create a set of potent and versatile dimerizers that can be used to control protein localization and interaction in vivo. The ability to control the biological potency of these dimerizers, as well as their ability to enter the cell, are important factors determining their usefulness for drug discovery applications.
    • Amara JF, Clackson T, Rivera VM, Guo T, Keenan T, Natesan S, Pollock R, Yang W, Courage NL, Holt DA, Gilman M. A versatile synthetic dimerizer for the regulation of protein - protein interactions. of special interest Proc Natl Acad Sci USA. 94:1997;10618-10623 Amara et al. describe the development of next generation synthetic dimerizers, based on ligands for FKBP-12. The ultimate aim is to create a set of potent and versatile dimerizers that can be used to control protein localization and interaction in vivo. The ability to control the biological potency of these dimerizers, as well as their ability to enter the cell, are important factors determining their usefulness for drug discovery applications.
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    • Amara, J.F.1    Clackson, T.2    Rivera, V.M.3    Guo, T.4    Keenan, T.5    Natesan, S.6    Pollock, R.7    Yang, W.8    Courage, N.L.9    Holt, D.A.10    Gilman, M.11
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    • of special interest of outstanding interest. This paper describes an RNA-based three-hybrid system which harnesses the power of the two-hybrid genetic screen and applies it to interactions of protein and RNA. A hybrid RNA molecule is used a simultaneously bind activation and binding domains fused to RNA-binding proteins. This new genetic screening method has been used to assay a AD - cDNA fusion library to isolate and clone genes whose protein products bind to specific RNA elements (see Martin et al. 1997 [25], Wang et al. 1996 [26] and Zhang et al. 1998 [28]) and to screen a library of hybrid RNAs for efficient binding to a known RNA-binding protein
    • SenGupta DJ, Zhang B, Kraemer B, Pochart P, Fields S, Wickens M. A three-hybrid system to detect RNA - protein interactions in vivo. of special interest of outstanding interest Proc Natl Acad Sci USA. 93:1996;8496-8501 This paper describes an RNA-based three-hybrid system which harnesses the power of the two-hybrid genetic screen and applies it to interactions of protein and RNA. A hybrid RNA molecule is used a simultaneously bind activation and binding domains fused to RNA-binding proteins. This new genetic screening method has been used to assay a AD - cDNA fusion library to isolate and clone genes whose protein products bind to specific RNA elements (see Martin et al. 1997 [25], Wang et al. 1996 [26] and Zhang et al. 1998 [28]) and to screen a library of hybrid RNAs for efficient binding to a known RNA-binding protein.
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    • Sengupta, D.J.1    Zhang, B.2    Kraemer, B.3    Pochart, P.4    Fields, S.5    Wickens, M.6
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    • The gene for histone RNA hairpin binding protein is located on human chromosome 4 and encodes a novel type of RNA binding protein
    • of special interest. The work described in this paper, along with Wang et al. 1996 [26], represent the first use of the three-hybrid system to screen an expression library for a protein that interacts with a specific target RNA. The target RNA was an element found in the 3′ end of histone mRNA which mediates pre-mRNA processing of the message, and which was known to bind stem-loop binding protein from diverse organisms. The gene encoding this protein was isolated from AD - cDNA libraries from humans, mice and frogs by the three-hybrid method.
    • Martin F, Schaller A, Eglite S, Schumperli D, Muller B. The gene for histone RNA hairpin binding protein is located on human chromosome 4 and encodes a novel type of RNA binding protein. of special interest EMBO J. 16:1997;769-778 The work described in this paper, along with Wang et al. 1996 [26], represent the first use of the three-hybrid system to screen an expression library for a protein that interacts with a specific target RNA. The target RNA was an element found in the 3′ end of histone mRNA which mediates pre-mRNA processing of the message, and which was known to bind stem-loop binding protein from diverse organisms. The gene encoding this protein was isolated from AD - cDNA libraries from humans, mice and frogs by the three-hybrid method.
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    • Martin, F.1    Schaller, A.2    Eglite, S.3    Schumperli, D.4    Muller, B.5
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    • The protein that binds the 3′ end of histone mRNA: A novel RNA-binding protein required for histone pre-mRNA processing
    • of special interest. See annotation [25].
    • Wang ZF, Whitfield ML, Ingledue TC 3rd, Dominski Z, Marzluff WF. The protein that binds the 3′ end of histone mRNA: a novel RNA-binding protein required for histone pre-mRNA processing. of special interest Genes Dev. 10:1996;3028-3040 See annotation [25].
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    • Wang, Z.F.1    Whitfield, M.L.2    Ingledue T.C. III3    Dominski, Z.4    Marzluff, W.F.5
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    • A conserved RNA-binding protein that regulates sexual fates in the C. elegans hermaphrodite germ line
    • of outstanding interest. Zhang et al. report the second successful cloning of an RNA-binding protein using the yeast three-hybrid system. Using an element from the 3′ UTR of the C. elegans fem-3 mRNA as bait, the authors isolated the gene encoding FBF, or fem-3 binding factor from an AD fusion library. This protein regulates the switch from spermatogenesis to oogenesis in C. elegans hermaphrodites through its interaction with the fem-3 mRNA. Particularly remarkable is the high efficiency with which the gene encoding this protein was isolated from the AD - cDNA library
    • Zhang B, Gallegos M, Puoti A, Durkin E, Fields S, Kimble J, Wickens MP. A conserved RNA-binding protein that regulates sexual fates in the C. elegans hermaphrodite germ line. of outstanding interest Nature. 1998; Zhang et al. report the second successful cloning of an RNA-binding protein using the yeast three-hybrid system. Using an element from the 3′ UTR of the C. elegans fem-3 mRNA as bait, the authors isolated the gene encoding FBF, or fem-3 binding factor from an AD fusion library. This protein regulates the switch from spermatogenesis to oogenesis in C. elegans hermaphrodites through its interaction with the fem-3 mRNA. Particularly remarkable is the high efficiency with which the gene encoding this protein was isolated from the AD - cDNA library.
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    • Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens
    • of outstanding interest. In this paper, Legrain's group describe an approach to the creation of a protein linkage map for the yeast S. cerevisiae. These workers screened a high-quality genomic library with small subsets of related proteins in the context of a DNA-binding domain fusion. Genes encoding probable in vivo parters for each bait were then chosen for second generation searches. In this manner, the nucleus of an interaction map for a set of yeast proteins involved in mRNA processing was created. Their approach will likely be of interest to those studying a particular aspect of yeast cell biology who wish to determine the list of players in a given cellular process.
    • Fromont-Racine M, Rain J-C, Legrain P. Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens. of outstanding interest Nat Genet. 16:1997;277-281 In this paper, Legrain's group describe an approach to the creation of a protein linkage map for the yeast S. cerevisiae. These workers screened a high-quality genomic library with small subsets of related proteins in the context of a DNA-binding domain fusion. Genes encoding probable in vivo parters for each bait were then chosen for second generation searches. In this manner, the nucleus of an interaction map for a set of yeast proteins involved in mRNA processing was created. Their approach will likely be of interest to those studying a particular aspect of yeast cell biology who wish to determine the list of players in a given cellular process.
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