UMMS Affiliation

Department of Microbiology and Physiological Systems

Publication Date

2018-03-28

Document Type

Article

Disciplines

Amino Acids, Peptides, and Proteins | Biochemical Phenomena, Metabolism, and Nutrition | Biochemistry, Biophysics, and Structural Biology | Cellular and Molecular Physiology | Enzymes and Coenzymes | Genetics and Genomics

Abstract

Stringent regulation of cellular levels of evolutionarily conserved centromeric histone H3 variant (CENP-A in humans, CID in flies, Cse4 in yeast) prevents its mislocalization to non-centromeric chromatin. Overexpression and mislocalization of CENP-A has been observed in cancers and leads to aneuploidy in yeast, flies, and human cells. Ubiquitin-mediated proteolysis of Cse4 by E3 ligases such as Psh1 and Sumo-Targeted Ubiquitin Ligase (STUbL) Slx5 prevent mislocalization of Cse4. Previously, we identified Siz1 and Siz2 as the major E3 ligases for sumoylation of Cse4. In this study, we have identified lysine 65 (K65) in Cse4 as a site that regulates sumoylation and ubiquitin-mediated proteolysis of Cse4 by Slx5. Strains expressing cse4 K65R exhibit reduced levels of sumoylated and ubiquitinated Cse4 in vivo Furthermore, co-immunoprecipitation experiments reveal reduced interaction of cse4 K65R with Slx5, leading to increased stability and mislocalization of cse4 K65R under normal physiological conditions. Based on the increased stability of cse4 K65R in psh1 strains but not in slx5 strains, we conclude that Slx5 targets sumoylated Cse4 K65 for ubiquitination-mediated proteolysis independent of Psh1. In summary, we have identified and characterized the physiological role of Cse4 K65 in sumoylation, ubiquitin-mediated proteolysis, and localization of Cse4 for genome stability.

Keywords

Cse4, E3 ubiquitin ligase, Kinetochore, Psh1, Saccharomyces cerevisiae, Slx5, Sumoylation, Ubiquitination

Rights and Permissions

Copyright © 2018 Ohkuni et al. G3 applies a Creative Commons Attribution License (CCAL) to all its published material. With this open access license, authors retain ownership to the copyright for their published article in G3. Authors allow anyone to share, remix, download, mine, or otherwise use their work-with the requirement that the authors and the journal are given the proper attribution (the full article citation). For more information, see https://creativecommons.org/licenses/by/4.0/. (per http://www.g3journal.org/content/scope-and-publication-policies#copyright)

DOI of Published Version

10.1534/g3.117.300419

Source

G3 (Bethesda). 2018 Mar 28;8(4):1215-1223. doi: 10.1534/g3.117.300419. Link to article on publisher's site

Journal/Book/Conference Title

G3 (Bethesda, Md.)

Related Resources

Link to Article in PubMed

PubMed ID

29432128

Creative Commons License

Creative Commons Attribution 4.0 License
This work is licensed under a Creative Commons Attribution 4.0 License.

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