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Related Experiment Videos

Using yeast to place human genes in functional categories.

Nianshu Zhang1, Michael Osborn, Paul Gitsham

  • 1School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Road, UK.

Gene
|February 1, 2003
PubMed
Summary

Researchers developed a yeast complementation system to identify human gene functions. This system successfully cloned human cDNAs that complement essential yeast genes, aiding in gene function discovery.

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Area of Science:

  • Genetics and Molecular Biology
  • Yeast Functional Genomics
  • Human Genome Annotation

Background:

  • The human genome project necessitates functional annotation of thousands of genes.
  • Model organisms like Saccharomyces cerevisiae are crucial for comparative genomics and functional studies.

Purpose of the Study:

  • To develop a robust system for cloning human cDNAs that functionally complement essential yeast genes.
  • To leverage Saccharomyces cerevisiae as a model eukaryote for human gene discovery.

Main Methods:

  • Development of a dual-promoter complementation system in Saccharomyces cerevisiae using tetO (doxycycline-repressible) and pMET3 (methionine-repressible) promoters.
  • Screening of human cDNA libraries for clones that restore growth of yeast strains with essential gene deletions under specific regulatory conditions.

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  • Validation of complementation by observing growth arrest upon repression of both yeast and human gene expression.

    • Main Results:

    • The system effectively identified functionally complementing human cDNAs from libraries.
    • Screening of 25 strains yielded human cDNAs complementing six essential yeast genes.
    • Discovery of a novel human ubiquitin-conjugating enzyme and a potential human signal peptidase.

    Conclusions:

    • The developed yeast complementation system is a powerful tool for functional genomics and human gene discovery.
    • This approach facilitates the identification of novel human genes and protein functions, including a PINT domain in human Psmd12.
    • Functional complementation in yeast provides a viable strategy for annotating the human genome.