Localization and degradation of Cln2 via the SCF Grr1 ubiquitin-proteosome pathway.

摘要

Cln2 is a G1 cyclin that associates with Cdk1 to promote the G1 to S transition in Saccharomyces cerevisiae. Previous studies have shown that Cln2 exhibits both nuclear and cytoplasmic localization and that localization may impart functionality. However, it is currently unknown what regulates Cln2 localization. The degradation of Cln2 is via phosphorylation-dependent SCFGrr1ubiquitin-mediated proteolysis. In this study, we explored the role of ubiquitin-mediated degradation in the localization of Cln2. A plasmid library of mutant CLN2 was created using gene synthesis and site-directed mutagenesis where lysines were mutated to arginines. Serial dilutions were used to assay for complementation on various media to determine whether various CLN2 mutants were viable. We successfully created a mutant plasmid library however the serial dilution assays were inconclusive. Thus at this time, no conclusion can be drawn about the functionality and localization of the CLN2 mutants.