Abstract
Cohesin mediates the 3-D structure of chromatin and is involved in maintaining genome stability and function. The cohesin core comprises Smc1 and Smc3, elongated-shaped proteins that dimerize through globular domains at their edges, called head and hinge. ATP binding to the Smc heads induces their dimerization and the formation of two active sites, while ATP hydrolysis results in head disengagement. This ATPase cycle is essential for driving cohesin activity. We report on the development of the first cohesin-inhibiting peptide (CIP). The CIP binds Smc3 in vitro and inhibits the ATPase activity of the holocomplex. Treating yeast cells with the CIP prevents cohesin's tethering activity and, interestingly, leads to the accumulation of cohesin on chromatin. CIP3 also affects cohesin activity in human cells. Altogether, we demonstrate the power of peptides to inhibit cohesin in cells and discuss the potential application of CIPs as a therapeutic approach.
| Original language | English |
|---|---|
| Article number | 107498 |
| Journal | iScience |
| Volume | 26 |
| Issue number | 9 |
| DOIs | |
| State | Published - 15 Sep 2023 |
Bibliographical note
Publisher Copyright:© 2023 The Authors
Funding
We thank Frank Uhlman for sharing the cohesin expression strains, Kobi Maman and Wessal Hanout for helping with the human cell line, Tamar Yulazery for advice on mitosis analysis in the human cell line experiments, and Michael Assa for his support with the Incucyte experiments. Thanks also to Steve Spencer for his professional editorial oversight. This work was funded by Israel Cancer Association grant #20221266 (IO) and Israel Science Foundation grant #987/19 (IO) and #401/18 (MD). Maria Elias performed this study as partial fulfillment of the requirements for a Ph.D. Degree in the Azrieli Faculty of Medicine, Bar-Ilan University. We thank Frank Uhlman for sharing the cohesin expression strains, Kobi Maman and Wessal Hanout for helping with the human cell line, Tamar Yulazery for advice on mitosis analysis in the human cell line experiments, and Michael Assa for his support with the Incucyte experiments. Thanks also to Steve Spencer for his professional editorial oversight. This work was funded by Israel Cancer Association grant #20221266 (IO) and Israel Science Foundation grant #987/19 (IO) and #401/18 (MD). M.E. and A.M. did the investigation. S.G. and Y.L. synthesized and purified peptides and supported FEB experiments and analysis. K.Y. analyzed chromosome condensation. C.T. cloned and set up the purification protocol of Smc3hd. A.P. and M.D. supported cohesin holocomplex purification. A.M. provided administrative support. N.Q. designed the peptides and supervised synthesis and purification. I.O. conceived the project, supervised experiments, and acquired funding. M.E. and A.V. prepared the figures M.E. A.M. N.Q. and I.O. wrote, reviewed, and edited the manuscript. S.G. Y.L. K.Y. C.T. A.P. and M.D. reviewed and commented on the manuscript. The authors read and approved the final manuscript. M.E. Y.L. S.G. N.Q. and I.O. applied for a patent for the molecules discussed in this paper (US provisional application number 63/383,706). All other authors declare no competing interests. We Support inclusive, diverse, and equitable conduct of research.
| Funders | Funder number |
|---|---|
| Azrieli Faculty of Medicine, Bar-Ilan University | |
| Israel Cancer Association | 20221266 |
| Israel Science Foundation | 401/18, 987/19 |
Keywords
- Cell biology
- Molecular biology
- Protein