Abstract
The cohesin complex plays an important role in the maintenance of genome stability. Cohesin is composed of four core subunits and a set of regulatory subunits that interact with the core subunits. Less is known about cohesin dynamics in live cells and on the contribution of individual subunits to the overall complex. Understanding the tethering mechanism of cohesin is still a challenge, especially because the proposed mechanisms are still not conclusive. Models proposed to describe tethering depend on either the monomeric cohesin ring or a cohesin dimer. Here, we investigate the role of cohesin dynamics and stoichiometry in live yeast cells at single-molecule resolution. We explore the effect of regulatory subunit deletion on cohesin mobility and found that depletion of different regulatory subunits has opposing effects. Finally, we show that cohesin exists mostly as a canonical monomer throughout the cell cycle, and its monomeric form is independent of its regulatory factors. Our results demonstrate that single-molecule tools have the potential to provide new insights into the cohesin mechanism of action in live cells.
| Original language | English |
|---|---|
| Article number | e48211 |
| Journal | EMBO Reports |
| Volume | 21 |
| Issue number | 2 |
| DOIs | |
| State | Published - 5 Feb 2020 |
Bibliographical note
Publisher Copyright:© 2019 The Authors
Funding
We thank Vinny Guaaci and Doug Koshland for providing yeast strains, and Ann L. Kirchmaier and Rong Li for providing PCH control yeast strains. W.L. thanks Jiji Chen and Brian D. Slaughter for helpful discussion on yeast FCS. W.L. thanks undergraduate students in J.I Laboratory. We also thank members of the Onn Laboratory for their support. This work was supported by the Israel Science Foundation Grant 1099/16 (IO). JI thanks the W.M. Keck Foundation for the support. We thank Vinny Guaaci and Doug Koshland for providing yeast strains, and Ann L. Kirchmaier and Rong Li for providing PCH control yeast strains. W.L. thanks Jiji Chen and Brian D. Slaughter for helpful discussion on yeast FCS. W.L. thanks undergraduate students in J.I Laboratory. We also thank members of the Onn Laboratory for their support. This work was supported by the Israel Science Foundation Grant 1099/16 (IO). JI thanks the W.M. Keck Foundation for the support.
| Funders | Funder number |
|---|---|
| Vinny Guaaci and Doug Koshland | |
| W. M. Keck Foundation | |
| Israel Science Foundation | 1099/16 |
Keywords
- SMC complexes
- chromosome
- cohesin
- fluorescence correlation spectroscopy
- photon counting histogram