An expanded tool kit for the auxin‐inducible degron system in budding yeast
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TLDR
The construction of a series of vectors is reported that significantly enhance the versatility of this auxin‐inducible degron (AID) system in Saccharomyces cerevisiae and provides evidence for a general usefulness of the system.Abstract:
Fusion of inducible degradation signals, so-called degrons, to cellular proteins is an elegant method of controlling protein levels in vivo. Recently, a degron system relying on the plant hormone auxin has been described for use in yeast and vertebrate cells. We now report the construction of a series of vectors that significantly enhance the versatility of this auxin-inducible degron (AID) system in Saccharomyces cerevisiae. We have minimized the size of the degron and appended a series of additional epitope tags, allowing detection by commercial antibodies or fluorescence microscopy. The vectors are compatible with PCR-based genomic tagging strategies, allow for C- or N-terminal fusion of the degron, and provide a range of selection markers. Application to a series of yeast proteins, including essential replication factors, provides evidence for a general usefulness of the system.read more
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Journal ArticleDOI
Slx5p-Slx8p Promotes Accurate Chromosome Segregation by Mediating the Degradation of Synaptonemal Complex Components during Meiosis.
Chao Liu,Haichao Zhao,Sai Xiao,Tingting Han,Yinghong Chen,Tong Wang,Yanjie Ma,Hui Gao,Zhiping Xie,Li-Lin Du,Jian Li,Guoping Li,Guoping Li,Wei Li +13 more
TL;DR: It is reported that a SUMO‐targeted ubiquitin ligase, Slx8p, promotes accurate chromosome segregation during meiosis, since the deletion of SLX8 leads to increased aneuploidy due to a defect in synaptonemal complex (SC) component degradation.
Journal ArticleDOI
A One-step strategy to target essential factors with auxin-inducible degron system in mouse embryonic stem cells
Jingsheng Li,Chunhong Dai,Wenyan Xie,Heyao Zhang,Xin Huang,Constantinos Chronis,Ying Ye,Wensheng Zhang +7 more
TL;DR: This work develops a one-step strategy to successfully target and repress the endogenous pluripotent genes in mouse ESCs and replace their expression with AID fused transgenes, providing an efficient way for employing the AID system to uncover novel function of essential pluripotency and chromatin modifier genes in ESCs.
Journal ArticleDOI
A toolbox for systematic discovery of stable and transient protein interactors in baker's yeast
TL;DR: Auxin-induced BiOtin LIgase diminiSHing (ABOLISH) was proposed in this article for downregulating endogenous biotinylation in yeast, where the endoplasmic reticulum insertase complex (EMC) was used to identify new substrates.
Journal ArticleDOI
Auxin-inducible degron 2 system deciphers functions of CTCF domains in transcriptional regulation
Judith Hyle,Mohamed Nadhir Djekidel,Justin Williams,Shaela Wright,Ying Shao,Beisi Xu,Chunliang Li +6 more
TL;DR: In this article , an improved auxin-inducible degron technology, AID2, was employed to facilitate the study of acute depletion of CTCF while overcoming the limitations of the previous AID system.
Posted ContentDOI
PICH translocase activity is required for proper distribution of SUMOylated proteins on mitotic chromosomes
Victoria A. Hassebroek,Hyewon Park,Nootan Pandey,Brooklyn T. Lerbakken,Vasilisa Aksenova,Alexei Arnaoutov,Mary Dasso,Yoshiaki Azuma +7 more
TL;DR: In this article, the authors show that PICH specifically promotes the redistribution of SUMOylated proteins like TopoisomeraseIIα (TopoIIα) on mitotic chromosomes.
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