Optimization of Sic1-Cln2-based phosphodegron tag for inducible protein degradation
Date
2021
Authors
Journal Title
Journal ISSN
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Publisher
Tartu Ülikool
Abstract
Precise regulation of protein abundance in the cell would be instrumental in both research
and industry. A variety of tools exists for the regulation of protein expression at the
transcriptional level. However, there is a shortage of tools that would allow control over
already expressed proteins. In this study, we exploit the native cell cycle regulation system
of Saccharomyces cerevisiae to develop a system for targeted inducible protein degradation.
Using sequence elements that modulate cyclin-dependent kinase (CDK) substrate
specificity, we generated a set of phosphodegron tags that have diverse effects on
degradation rates of the tagged proteins. The control over the system is provided by
conditional expression of the stabilized version of cyclin Clb3, which drives CDK-dependent
phosphorylation of the tag, and F-box protein Grr1, which recognizes phosphorylated tag
and label it for further degradation. In the course of this work, we have designed a set of
phosphodegron tags that can be used for the regulation of the protein abundance in the cell
and its maintenance at desirable levels that can be useful for the industrial production of
value-added products. In addition, we discovered that Cks1 priming effect on the multisite
phosphorylation is not only distance-, but, likely, also context-dependent which might be of
interest for fundamental research.
Description
Keywords
cell cycle, degrons, cyclins, Sic1