Novel methods to study genes with multiple functions


Novel methods to study genes with multiple functions



To establish a method for conditional disruption of protein function in Caenorhabditis elegans, we are implementing system in which a degron tag is added to a protein of interest. Under certain conditions the tagged protein is recognized by a ubiquitin ligase that catalyzes polyubiquitination of the tag and degradation of the protein. Degron tags are commonly used in yeast and cultured cells. However, the standard degrons cannot be used in C. elegans because the restrictive conditions are lethal to worms. Recently, an Auxin Inducible Degron (AID) has been shown to rapidly reduce protein levels in budding yeast, fission yeast, and cells derived from mice, hamsters, chickens, and humans. AID works by tagging the target protein with a polypeptide from plants. In the presence of the hormone auxin, the polypeptide tag is bound by the plant protein TIR1. TIR1 in turn recruits several of the cell’s own components of the ubiquitin ligase complex, resulting in polyubiquitination and degradation of the tagged protein. Importantly, proteins tagged with the AID degron function normally until exposed to auxin, and degradation of AID-tagged proteins occurs in conditions compatible with growth of C. elegans strains.

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Novel methods to study genes with multiple functions