PT  - JOURNAL ARTICLE
AU  - Lucía Durrieu
AU  - Rikard Johansson
AU  - Alan Bush
AU  - David Janzén
AU  - Martin Gollvik
AU  - Gunnar Cedersund
AU  - Alejandro Colman-Lerner
TI  - Quantification of nuclear transport in single cells
AID  - 10.1101/001768
DP  - 2014 Jan 01
TA  - bioRxiv
PG  - 001768
4099  - http://biorxiv.org/content/early/2014/01/11/001768.short
4100  - http://biorxiv.org/content/early/2014/01/11/001768.full
AB  - Regulation of nuclear transport is a key cellular function involved in many central processes, such as gene expression regulation and signal transduction. Rates of protein movement between cellular compartments can be measured by FRAP. However, no standard and reliable methods to calculate transport rates exist. Here we introduce a method to extract import and export rates, suitable for noisy single cell data. This method consists of microscope procedures, routines for data processing, an ODE model to fit to the data, and algorithms for parameter optimization and error estimation.Using this method, we successfully measured import and export rates in individual yeast. For YFP, average transport rates were 0.15 sec-1. We estimated confidence intervals for these parameters through likelihood profile analysis. We found large cell-to-cell variation (CV = 0.79) in these rates, suggesting a hitherto unknown source of cellular heterogeneity. Given the passive nature of YFP diffusion, we attribute this variation to large differences among cells in the number or quality of nuclear pores.Owing to its broad applicability and sensitivity, this method will allow deeper mechanistic insight into nuclear transport processes and into the largely unstudied cell-to-cell variation in kinetic rates.