Characterization of cell-to-cell variation in nuclear transport rates and identification of its sources

Nuclear transport is an essential part of eukaryotic cell function. Here, we present scFRAP, a model-assisted fluorescent recovery after photobleaching (FRAP)- based method to determine nuclear import and export rates independently in individual live cells. To overcome the inherent noise of single-cell measurements, we performed sequential FRAPs on the same cell. We found large cell-to-cell variation in transport rates within isogenic yeast populations. For passive transport, the variability in NPC number might explain most of the variability. Using this approach, we studied mother-daughter cell asymmetry in the active nuclear shuttling of the transcription factor Ace2, which is specifically concentrated in daughter cell nuclei in early G1. Rather than reduced export in the daughter cell, as previously hypothesized, we found that this asymmetry is mainly due to an increased import in daughters. These results shed light on cell-to-cell variation in cellular dynamics and its sources. [Display omitted] •Sequential FRAPs allow precise estimation of nuclear shuttling rates in single cells•There is high cell-to-cell variation in nuclear import and export rates•Most cell-to-cell variation stems from variability in core transport machinery•Ace2 daughter-cell enrichment is due to higher import rates in that cell type Biological sciences; Cell biology; Mathematical biosciences