A zygote-based assay to evaluate intranuclear shuttling in S. cerevisiae

It is often necessary to learn whether macromolecules occupy a fixed place in cells. This protocol makes it possible to learn whether individual nucleolar proteins in S. cerevisiae remain in place or depart from and return to the nucleolus. The protocol uses early zygotes in which parental nucleoli are separate for at least one hour. The protocol demonstrates that the localization of many nucleolar proteins is in fact highly dynamic. Photobleaching is not required. For complete details on the use and execution of this protocol, please refer to Tartakoff et al. (2021).