Dynamic proteomics in individual human cells uncovers widespread cell-cycle dependence of nuclear proteins.
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.
Sigal
Alex
A
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot 76100, Israel.
Milo
Ron
R
Cohen
Ariel
A
Geva-Zatorsky
Naama
N
Klein
Yael
Y
Alaluf
Inbal
I
Swerdlin
Naamah
N
Perzov
Natalie
N
Danon
Tamar
T
Liron
Yuvalal
Y
Raveh
Tal
T
Carpenter
Anne E
AE
Lahav
Galit
G
Alon
Uri
U
D001426:Bacterial Proteins / Q000737:chemistry; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000502:physiology*; D002999:Clone Cells; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods; D008164:Luminescent Proteins / Q000737:chemistry; D009687:Nuclear Proteins / Q000737:chemistry* / Q000502:physiology*; D019151:Peptide Library; D040901:Proteomics
D001426:Bacterial Proteins; D018797:Cell Cycle Proteins; D008164:Luminescent Proteins; D009687:Nuclear Proteins; D019151:Peptide Library; C054562:yellow fluorescent protein, Bacteria