Characterization of BioID tagging systems in budding yeast and exploring the interactome of the Ccr4-Not complex.

ABSTRACT: The modified Escherichia coli biotin ligase BirA* was the first developed for proximity labeling of proteins (BioID). However, it has low activity at temperatures below 37°C, which reduces its effectiveness in organisms growing at lower temperatures, such as budding yeast. Multiple derivatives of the enzymes have been engineered, but a thorough comparison of these variations of biotin ligases and the development of versatile tools for conducting these experiments in Saccharomyces cerevisiae would benefit the community. Here, we designed a suite of vectors to compare the activities of biotin ligase enzymes in yeast. We found that the newer TurboID versions were the most effective at labeling proteins, but they displayed low constitutive labeling of proteins even in the absence of exogenous

SUBMITTER: Pfannenstein J 

PROVIDER: S-EPMC11540327 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature