YEAST MRNA 3' END FORMATION AS AN ANTIFUNGAL DRUG TARGET

THE FORMATION OF POLYADENYLATED 3'-TERMINI IS AN ESSENTIAL STEP IN THE BIOGENESIS OF EUKARYOTIC MRNA. IN VERTEBRATES, THE CONSENSUS AAUAAA AND OTHER SEQUENCES HAVE BEEN IDENTIFIED AS REQUIREMENTS FOR CORRECT 3'-END FORMATION. IN YEAST, ONLY HALF OF ALL GENES HAVE A RECOGNIZABLE AAUAAA CONSENSUS, AND THIS PROCESS APPEARS TO BE SUBSTANTIALLY DIFFERENT. THE SPECIFIC AIMS IN PHASE I ARE TO (1) CONSTRUCT YEAST EXPRESSION VECTORS THAT EXPRESS ONLY WHEN 3'-END FORMATION IS INHIBITED, AND (2) CONFIRM THAT SACCHAROMYCES CEREVISIAE WILL EXPRESS OTHER FUNGAL GENES BY PROCESSING 3'-ENDS CORRECTLY. THE LONG-TERM GOALS IN PHASE II WILL BE TO (1) IDENTIFY AND CLONE GENES THAT PROCESS 3'-ENDS, AND (2) FURTHER TEST AND DEVELOP AN IN VITRO SYSTEM FOR 3'-END FORMATION.