Inducible protein degradation in Saccharomyces cerevisiae

Abstract

Recently, several synthetic biology toolboxes have been developed to be used in S.cerevisiae. The application of such toolbox components helps to improve bioproduction of desired chemicals or heterologous enzymes. In this work, we designed a Clb3-CDC28 (cyclin-cyclin dependent kinase) specific degron module that can be used as a synthetic biology tool in order to direct the metabolic dynamics of S.cerevisiae. The results showed that the degradation of such degron module conjugated proteins are mediated by the phosphorylation of the CDC28 target residues at Cln2 degron domain and this phosphorylation is predominantly carried out by Clb3-CDC28 complex which makes the degradation controllable. However, we also showed that expression of Clb3-CDC28 is not the only factor that drives the degradation of degron module conjugated proteins by leaving an empty room to improve the controllability of the system. In Estonian: Pagaripärmi jaoks on viimasel ajal arendatud mitmeid sünteetilise bioloogia tööriistakaste. Taoliste tööriistakastide abil saab rakkudes toota mitmeid vajalikke kemikaale ja ensüüme. Bakalaureuse töö käigus disainiti Clb3-CDC28 spetsiifiline degronimoodul, mille abil saab mõjutada rakkude metabolismi S. cerevisea’s. Töö tulemusel selgus, et Clb3-CDC28 spetsiifilise fosforüülimisega saab mõjutada Cln2 degronil põhineva mooduliga seotud valkude tasemeid. Samas selgus, et degronimooduli lisamine valkudele mõjutab nende taset sõltumata Clb3-CDC28 indutseerimisest, mistõttu tuleb tulevikus antud süsteemi täiustada.