Access

Article

Nature Methods 3, 525–531 (1 July 2006) | doi:10.1038/nmeth892

Dynamic proteomics in individual human cells uncovers widespread cell-cycle dependence of nuclear proteins

Alex Sigal , Ron Milo , Ariel Cohen , Naama Geva-Zatorsky , Yael Klein , Inbal Alaluf , Naamah Swerdlin , Natalie Perzov , Tamar Danon , Yuvalal Liron , Tal Raveh , Anne E Carpenter , Galit Lahav & Uri Alon

We examined cell cycle–dependent changes in the proteome of human cells by systematically measuring protein dynamics in individual living cells. We used time-lapse microscopy to measure the dynamics of a random subset of 20 nuclear proteins, each tagged with yellow fluorescent protein (YFP) at its endogenous chromosomal location. We synchronized the cells in silico by aligning protein dynamics in each cell between consecutive divisions. We observed widespread (40%) cell-cycle dependence of nuclear protein levels and detected previously unknown cell cycle–dependent localization changes. This approach to dynamic proteomics can aid in discovery and accurate quantification of the extensive regulation of protein concentration and localization in individual living cells.