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

Research output: Contribution to journalArticlepeer-review

112 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)525-531
Number of pages7
JournalNature Methods
Volume3
Issue number7
DOIs
StatePublished - Jul 2006
Externally publishedYes

Bibliographical note

Funding Information:
We thank Quantomix for plasmid construction; Z. Kam for microscopy guidance; B. Zimmerman for cytoskeleton staining; E. Ariel and A. Sharp from the Weizmann flow cytometry unit for assistance, and M. Springer, P. Bordalo, O. Zuk and members of the Alon lab for discussions of the manuscript. We thank the Kahn Family Foundation and the Israel Science Foundation for support. R.M. thanks the Horowitz Complexity Science Foundation for support. A.E.C. thanks the Novartis Life Science Foundation for support.

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