scholarly journals Yeast Two-Hybrid Systems and Protein Interaction Mapping Projects for Yeast and Worm

Yeast ◽  
2000 ◽  
Vol 1 (2) ◽  
pp. 88-94 ◽  
Author(s):  
Albertha J. M. Walhout ◽  
Simon J. Boulton ◽  
Marc Vidal
Keyword(s):  
Hybrid Systems ◽  
High Throughput ◽  
Protein Interaction ◽  
Protein Interactions ◽  
Large Scale ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Systematic Analysis ◽  
Technical Features ◽  
Two Hybrid

The availability of complete genome sequences necessitates the development of standardized functional assays to analyse the tens of thousands of predicted gene products in high-throughput experimental settings. Such approaches are collectively referred to as ‘functional genomics’. One approach to investigate the properties of a proteome of interest is by systematic analysis of protein–protein interactions. So far, the yeast two-hybrid system is the most commonly used method for large-scale, high-throughput identification of potential protein–protein interactions. Here, we discuss several technical features of variants of the two-hybrid systems in light of data recently obtained from different protein interaction mapping projects for the budding yeast Saccharomyces cerevisiae and the nematode Caenorhabditis elegans.

scholarly journals Yeast Two-Hybrid Systems and Protein Interaction Mapping Projects for Yeast and Worm

Yeast ◽  
2000 ◽  
Vol 1 (2) ◽  
pp. 88-94 ◽  
Author(s):  
Albertha J. M. Walhout ◽  
Simon J. Boulton ◽  
Marc Vidal
Keyword(s):  
Hybrid Systems ◽  
High Throughput ◽  
Protein Interaction ◽  
Protein Interactions ◽  
Large Scale ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Systematic Analysis ◽  
Technical Features ◽  
Two Hybrid

The availability of complete genome sequences necessitates the development of standardized functional assays to analyse the tens of thousands of predicted gene products in high-throughput experimental settings. Such approaches are collectively referred to as ‘functional genomics’. One approach to investigate the properties of a proteome of interest is by systematic analysis of protein–protein interactions. So far, the yeast two-hybrid system is the most commonly used method for large-scale, high-throughput identification of potential protein–protein interactions. Here, we discuss several technical features of variants of the two-hybrid systems in light of data recently obtained from different protein interaction mapping projects for the budding yeastSaccharomyces cerevisiaeand the nematodeCaenorhabditis elegans.

scholarly journals High throughput flow cytometry based yeast two-hybrid array approach for large-scale analysis of protein-protein interactions

2011 ◽  
Vol 81A (1) ◽  
pp. 90-98 ◽  
Author(s):  
Jun Chen ◽  
Mark B. Carter ◽  
Bruce S. Edwards ◽  
Hong Cai ◽  
Larry A. Sklar
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Protein Interactions ◽  
Large Scale ◽  
Scale Analysis ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Large Scale Analysis ◽  
Two Hybrid

On the structure of protein–protein interaction networks

2003 ◽  
Vol 31 (6) ◽  
pp. 1491-1496 ◽  
Author(s):  
A. Thomas ◽  
R. Cannings ◽  
N.A.M. Monk ◽  
C. Cannings
Keyword(s):  
Power Law ◽  
Protein Interactions ◽  
Large Scale ◽  
Underlying Structure ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Protein Protein Interaction ◽  
Human Proteins ◽  
Approximate Power ◽  
Two Hybrid

We present a simple model for the underlying structure of protein–protein pairwise interaction graphs that is based on the way in which proteins attach to each other in experiments such as yeast two-hybrid assays. We show that data on the interactions of human proteins lend support to this model. The frequency of the number of connections per protein under this model does not follow a power law, in contrast to the reported behaviour of data from large-scale yeast two-hybrid screens of yeast protein–protein interactions. Sampling sub-graphs from the underlying graphs generated with our model, in a way analogous to the sampling performed in large-scale yeast two-hybrid searches, gives degree distributions that differ subtly from the power law and that fit the observed data better than the power law itself. Our results show that the observation of approximate power law behaviour in a sampled sub-graph does not imply that the underlying graph follows a power law.

High-Throughput Yeast Two-Hybrid Assays for Large-Scale Protein Interaction Mapping

Methods ◽  
2001 ◽  
Vol 24 (3) ◽  
pp. 297-306 ◽  
Author(s):  
Albertha J.M. Walhout ◽  
Marc Vidal
Keyword(s):  
High Throughput ◽  
Protein Interaction ◽  
Large Scale ◽  
Yeast Two Hybrid ◽  
Two Hybrid

scholarly journals A High-ThroughputCandida albicansTwo-Hybrid System

mSphere ◽  
2018 ◽  
Vol 3 (4) ◽  
Author(s):  
Floris Schoeters ◽  
Carol A. Munro ◽  
Christophe d’Enfert ◽  
Patrick Van Dijck
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Candida Albicans ◽  
High Throughput ◽  
Protein Interactions ◽  
Fungal Pathogen ◽  
Model Organism ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Content Type ◽  
Two Hybrid

ABSTRACTCandida albicansis a human fungal pathogen that does not follow the universal codon usage, as it translates the CUG codon into serine rather than leucine. This makes it difficult to study protein-protein interactions using the standard yeast two-hybrid (Y2H) system in the model organismSaccharomyces cerevisiae. Due to the lack of adapted tools, only a small number of protein-protein interactions (PPIs) have been detected or studied usingC. albicans-optimized tools despite the importance of PPIs to understand cell biology. However, with the sequencing of the whole genome ofC. albicans, the availability of an ORFeome collection containing 5,099 open reading frames (ORFs) in Gateway-adapted donor vectors, and the creation of a Gateway-compatibleC. albicans-specific two-hybrid (C2H) system, it became possible to study protein-protein interactions on a larger scale usingC. albicansitself as the model organism. Erroneous translations are hereby eliminated compared to using theS. cerevisiaeY2H system. Here, we describe the technical adaptations and the first application of the C2H system for a high-throughput screen, thus making it possible to screen thousands of PPIs at once inC. albicansitself. This first, small-scale high-throughput screen, using Pho85 as a bait protein against 1,646 random prey proteins, yielded one interacting partner (Pcl5). The interaction found with the high-throughput setup was further confirmed with a low-throughput C2H experiment and with a coimmunoprecipitation (co-IP) experiment.IMPORTANCECandida albicansis a major fungal pathogen, and due to the rise of fungal infections and emerging resistance to the limited antifungals available, it is important to develop novel and more specific antifungals. Protein-protein interactions (PPIs) can be applied as very specific drug targets. However, because of the aberrant codon usage ofC. albicans, the traditional yeast two-hybrid system inSaccharomyces cerevisiaeis difficult to use, and only a limited number of PPIs have been described inC. albicans. To overcome this, aC. albicanstwo-hybrid (C2H) system was developed in 2010. The current work describes, for the first time, the application of the C2H system in a high-throughput setup. We hereby show the usefulness of the C2H system to investigate and detect PPIs inC. albicans, making it possible to further elucidate protein networks inC. albicans, which has the potential to lead to the development of novel antifungals which specifically disrupt PPIs important for virulence.

scholarly journals Unraveling protein interactions between the temperate virus Bam35 and its Bacillus host using an integrative yeast two hybrid-high throughput sequencing approach

2021 ◽  
Author(s):  
Ana Lechuga ◽  
Cédric Lood ◽  
Mónica Berjón-Otero ◽  
Alicia Del Prado ◽  
Jeroen Wagemans ◽  
...  
Keyword(s):  
High Throughput ◽  
Protein Interactions ◽  
High Throughput Sequencing ◽  
Genomic Library ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Protein Protein Interaction ◽  
Novel Approach ◽  
Diverse Groups ◽  
Two Hybrid

Bacillus virus Bam35 is the model Betatectivirus and member of the Tectiviridae family, which is composed of tailless, icosahedral, and membrane-containing bacteriophages. The interest in these viruses has greatly increased in recent years as they are thought to be an evolutionary link between diverse groups of prokaryotic and eukaryotic viruses. Additionally, betatectiviruses infect bacteria of the Bacillus cereus group, known for their applications in industry and notorious since it contains many pathogens. Here, we present the first protein-protein interactions network for a tectivirus-host system by studying the Bam35- Bacillus thuringiensis model using a novel approach that integrates the traditional yeast two-hybrid system and Illumina high-throughput sequencing. We generated and thoroughly analyzed a genomic library of Bam35’s host B. thuringiensis HER1410 and screened interactions with all the viral proteins using different combinations of bait-prey couples. In total, this screen resulted in the detection of over 4,000 potential interactions, of which 183 high-confidence interactions were defined as part of the core virus-host interactome. Overall, host metabolism proteins and peptidases are particularly enriched within the detected interactions, distinguishing this host-phage system from the other reported host-phage protein-protein interaction networks (PPIs). Our approach also suggests biological roles for several Bam35 proteins of unknown function, resulting in a better understanding of the Bam35- B. thuringiensis interaction at the molecular level.

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