Nature Methods 3, 525 - 531 (2006)
Published online: 21 June 2006; | doi:10.1038/nmeth892
Dynamic proteomics in individual human cells uncovers widespread cell-cycle dependence of nuclear proteinsAlex Sigal1, 6, Ron Milo1, 2, 6, Ariel Cohen1, Naama Geva-Zatorsky1, Yael Klein1, Inbal Alaluf1, Naamah Swerdlin1, Natalie Perzov1, Tamar Danon1, Yuvalal Liron1, Tal Raveh3, Anne E Carpenter4, Galit Lahav5
& Uri Alon1, 21
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel. 2
Department of Physics of Complex Systems, Weizmann Institute of Science, Rehovot 76100, Israel. 3
Department of Developmental Biology, Stanford University, Palo Alto, California 94305-5439, USA. 4
Whitehead Institute for Biomedical Research, Cambridge, Massachusetts 02142, USA. 5
Department of Systems Biology, Harvard Medical School, Boston, Massachusetts 02115, USA. 6
These authors contributed equally to this work.
Correspondence should be addressed to Uri Alon uri.alon@weizmann.ac.il 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.
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, 0.7 numerical aperture (NA) phase objective and a filter cube containing a filter set for YFP (Chroma)
