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Y2H System

Yeast 2 hybrid system is a type of molecular biology technique for mapping protein protein interactions

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05 - JANVI BHATANKAR
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53 views12 pages

Y2H System

Yeast 2 hybrid system is a type of molecular biology technique for mapping protein protein interactions

Uploaded by

05 - JANVI BHATANKAR
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PPTX, PDF, TXT or read online on Scribd
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The Yeast Two-Hybrid

System: A Tool for Mapping


Protein–Protein Interactions

Janvi Bhatankar
Roll no.2
Contents
• Introduction to Protein-Protein Interactions
• Overview of the Yeast Two-Hybrid System
• Rationale for the Yeast Two-Hybrid System
• Components of the Y2H System
• Advantages of Library Screening
• Critical Parameters in Y2H
• Evaluation of Y2H Results
• Applications of Y2H
• Conclusion
• Reference
Introduction to Protein-Protein Interactions
• PPIs as Cellular Regulators: Protein interactions regulate cellular functions.
• Role in Disease Mechanisms: PPIs are key in diseases like cancer and viral
infections.
• Complexity of Networks: Millions of PPIs are required for normal cellular
operations.
Overview of the Yeast Two-Hybrid System
• Purpose: Identifies and maps protein-protein interactions (PPIs).

• Mechanism: Uses reporter gene activation to detect interactions.

• High-Throughput Capability: Can be applied for genome-wide

screening.
Rationale for the Yeast Two-Hybrid System
• Reconstitution of Transcription Factor: Interaction reconstitutes a transcription factor.

• DNA Binding and Activation Domains: Split domains fuse with target proteins (bait and prey).

• Reporter Gene Activation: Interaction leads to the expression of reporter genes (e.g., HIS3).
Components of the Y2H System
• Bait Protein: Protein X fused to the DNA-binding domain (DBD).

• Prey Protein: Protein fragments fused to the activation domain (AD).

• Reporter Genes: Activated reporter genes indicate successful PPIs.


Critical Parameters in Y2H
• Vector Choice: Different vectors for DBD and AD fusions affect results.

• Self-Activation Check: Baits may activate reporter genes without prey interaction.

• Strain Compatibility: Bait and prey strains must be compatible for mating.
Evaluation of Y2H Results
• Growth Comparison: Interacting pairs grow; non-interacting pairs do not.

• Filtering False Positives: "Sticky" preys may produce false interactions.

• Retesting: Retesting positive interactions improves result reliability.


Applications of Y2H
• Proteome Mapping: Used for mapping protein interaction networks.

• Disease Research: Applied to study cancer, viral infections, and other diseases.

• Functional Genomics: Helps in annotating gene functions across organisms.


Conclusion
• Key Role in Proteomics: Y2H remains a vital tool in mapping PPIs.

• Ongoing Innovations: Constant improvements are increasing efficiency and accuracy.

• Broad Applicability: Applicable across diverse biological fields, from basic science to

medical research.
Reference:
• Lin JS, Lai EM. Protein-Protein Interactions: Yeast Two-Hybrid System.
Methods Mol Biol. 2017;1615:177-187. doi: 10.1007/978-1-4939-7033-
9_14. PMID: 28667612.
Thank you

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