IDENTIFICATION AND USE OF COMPOUNDS THAT AFFECT THE FIDELITY OF EUKARYOTIC TRANSLATION INITIATION CODON SELECTION

摘要

A screening method for identifying compounds that alter the fidelity with which the initiation codon in mRNAs is recognized by the translational apparatus in eukaryotes is disclosed. This screening method was used to identify compounds having such activity. Methods of altering the fidelity of initiation codon selection are also disclosed. Methods of treating disorders characterized by single nucleotide mutations in initiation codons using compounds identified by the screening method, as well as methods of treating fungal and parasitic infections and hyperproliferative disorders using compounds identified by the screening method are also disclosed.

主权项

1. A method for identifying an active compound affecting fidelity of eukaryotic translation initiation codon selection comprising:
introducing a compound to eukaryotic cells in culture, wherein the cells comprise a DNA sequence encoding a first reporter protein, and the mRNA from the first reporter protein has an initiation codon that is a near-cognate of AUG; measuring an amount of the first reporter protein; determining a change in the amount of first reporter protein by comparing the amount of the first reporter protein to a baseline amount of the first reporter protein measured in the absence of the compound; and selecting the compound as the active compound for affecting fidelity of eukaryotic translation initiation codon of the near-cognate of AUG initiation codons when the change in the amount of the first reporter protein meets or exceeds a predetermined threshold.