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

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

Research output: Contribution to journal › Article › peer-review

115 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 language	English
Pages (from-to)	525-531
Number of pages	7
Journal	Nature Methods
Volume	3
Issue number	7
DOIs	https://doi.org/10.1038/nmeth892
State	Published - Jul 2006
Externally published	Yes

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.

Funding

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.

Funders
Horowitz Complexity Science Foundation
Kahn Family Foundation
Novartis Life Science Foundation
Israel Science Foundation

Access to Document

10.1038/nmeth892

Cite this

@article{effeaa31c78746cba55b302f8860af52,

title = "Dynamic proteomics in individual human cells uncovers widespread cell-cycle dependence of nuclear proteins",

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.",

author = "Alex Sigal and Ron Milo and Ariel Cohen and Naama Geva-Zatorsky and Yael Klein and Inbal Alaluf and Naamah Swerdlin and Natalie Perzov and Tamar Danon and Yuvalal Liron and Tal Raveh and Carpenter, \{Anne E.\} and Galit Lahav and Uri Alon",

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.",

year = "2006",

month = jul,

doi = "10.1038/nmeth892",

language = "אנגלית",

volume = "3",

pages = "525--531",

journal = "Nature Methods",

issn = "1548-7091",

publisher = "Nature Research",

number = "7",

}

TY - JOUR

T1 - Dynamic proteomics in individual human cells uncovers widespread cell-cycle dependence of nuclear proteins

AU - Sigal, Alex

AU - Milo, Ron

AU - Cohen, Ariel

AU - Geva-Zatorsky, Naama

AU - Klein, Yael

AU - Alaluf, Inbal

AU - Swerdlin, Naamah

AU - Perzov, Natalie

AU - Danon, Tamar

AU - Liron, Yuvalal

AU - Raveh, Tal

AU - Carpenter, Anne E.

AU - Lahav, Galit

AU - Alon, Uri

N1 - 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.

PY - 2006/7

Y1 - 2006/7

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/33745391854

U2 - 10.1038/nmeth892

DO - 10.1038/nmeth892

M3 - ???researchoutput.researchoutputtypes.contributiontojournal.article???

SN - 1548-7091

VL - 3

SP - 525

EP - 531

JO - Nature Methods

JF - Nature Methods

IS - 7

ER -

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

Abstract

Bibliographical note

Funding

Access to Document

Other files and links

Fingerprint

Cite this