High-sensitivity measurements of multiple kinase activities in live single cells

Regot, Sergi; Hughey, Jacob J.; Bajar, Bryce T.; Carrasco, Silvia; Covert, Markus W.

doi:10.1016/j.cell.2014.04.039

Back to Previous Page

- Add this to...
- Twitter
- Facebook
- Google Bookmarks
- LinkedIn

High-sensitivity measurements of multiple kinase activities in live single cells

Published Date:

Jun 19 2014

Publisher's site:

http://dx.doi.org/10.1016/j.cell.2014.04.039 New Site Icon

Source:

Cell. 2014; 157(7):1724-1734.

[PDF-3.43 MB]

Details:

Personal Authors:

Regot, Sergi ; Hughey, Jacob J. ; Bajar, Bryce T. ; Carrasco, Silvia ; Covert, Markus W. ;

Keywords:

[+]

Pubmed ID:

24949979

Pubmed Central ID:

PMC4097317

Description:

Increasing evidence has shown that population dynamics are qualitatively different from single-cell behaviors. Reporters to probe dynamic, single-cell behaviors are desirable yet relatively scarce. Here, we describe an easy-to-implement and generalizable technology to generate reporters of kinase activity for individual cells. Our technology converts phosphorylation into a nucleocytoplasmic shuttling event that can be measured by epifluorescence microscopy. Our reporters reproduce kinase activity for multiple types of kinases and allow for calculation of active kinase concentrations via a mathematical model. Using this technology, we made several experimental observations that had previously been technicallyunfeasible, including stimulus-dependent patterns of c-Jun N-terminal kinase (JNK) and nuclear factor kappa B (NF-κB) activation. We also measured JNK, p38, and ERK activities simultaneously, finding that p38 regulates the peak number, but not the intensity, of ERK fluctuations. Our approach opens the possibility of analyzing a wide range of kinase-mediated processes in individual cells.

Document Type:

Journal Article

Collection(s):

CDC Public Access

Funding:

5DP1LM01150-05/DP/NCCDPHP CDC HHS/United States
DP1 LM011510/LM/NLM NIH HHS/United States
DP1 OD006413/OD/NIH HHS/United States
P50 GM107615/GM/NIGMS NIH HHS/United States
P50GM107615/GM/NIGMS NIH HHS/United States
R21 5R21AI104305-02/AI/NIAID NIH HHS/United States
R21 AI104305/AI/NIAID NIH HHS/United States

Supporting Files:

	NIHMS600454-supplement-01.pdf	application/pdf
	NIHMS600454-supplement-02.avi	application/x-troff-msvideo
	NIHMS600454-supplement-03.pdf	application/pdf
	nihms600454.nxml	application/octet-stream
	nihms600454f1.gif	image/gif
	nihms600454f1.jpg	image/jpeg
	nihms600454f2.gif	image/gif
	nihms600454f2.jpg	image/jpeg

No Related Documents.

You May Also Like:

Trajectories of cell-cycle progression from fixed cell populations

Personal Author:

Gut, Gabriele; Tadmor, Michelle D; Pe’er, Dana; Pelkmans, Lucas; Liberali, Prisca;

Published:

Aug 24 2015

Source:

Nat Methods. 12(10):951-954.

Description:

An accurate dissection of sources of cell-to-cell variability is crucial for quantitative biology at the single-cell level but has been challenging for the cell cycle. We present Cycler, a robust method that constructs a continuous trajectory of cell...

File Type:

[PDF - 731.54 KB]
Environmental assessment and exposure reduction of cockroaches: A practice parameter

Personal Author:

Portnoy, Jay; Chew, Ginger L.; Phipatanakul, Wanda; Williams, P. Brock; Grimes, Carl; Kennedy, Kevin; Matsui, Elizabeth C.; Miller, J. David; Bernstein, David; Blessing-Moore, Joann; Cox, Linda; Khan, David; Lang, David; Nicklas, Richard; Oppenheimer, John; Randolph, Christopher; Schuller, Diane; Spector, Sheldon; Tilles, Stephen A.; Wallace, Dana; Seltzer, James; Sublett, James;

Published:

Aug 09 2013

Source:

J Allergy Clin Immunol. 132(4).

Description:

This parameter was developed by the Joint Task Force on Practice Parameters, representing the American Academy of Allergy, Asthma & Immunology (AAAAI); the American College of Allergy, Asthma & Immunology (ACAAI); and the Joint Council of Allergy, As...

File Type:

[PDF - 164.81 KB]
ERK Oscillation-Dependent Gene Expression Patterns and Deregulation by Stress Response

Personal Author:

Waters, Katrina M.; Cummings, Brian S.; Shankaran, Harish; Scholpa, Natalie E.; Weber, Thomas J.;

Published:

Aug 12 2014

Source:

Chem Res Toxicol. 2014; 27(9):1496-1503.

Description:

Studies were undertaken to determine whether extracellular signal regulated kinase (ERK) oscillations regulate a unique subset of genes in human keratinocytes and subsequently whether the p38 stress response inhibits ERK oscillations. A DNA microarra...

File Type:

[PDF - 1.88 MB]
Applying public health strategies to primary immunodeficiency diseases; a potential approach to genetic disorders

Personal Author:

Baker, Timothy G.; Chen, Bin; Grosse, Scott D.; Hannon, W. Harry; Kalman, Lisa V.; Khoury, Muin J.; Kobrynski, Lisa; Lindegren, Mary Lou; Mattson, Marifran; McDougal, J. Steven; Moore, Cynthia A.; Rasmussen, Sonja A.; Spira, Thomas J.; Vanderford, Marsha L.; Vogt, Robert F.;

Corporate Author:

Centers for Disease Control and Prevention (U.S.); Centers for Disease Control and Prevention (U.S.), Public Health Practice Program Office., Division of Laboratory Systems.; National Center for Environmental Health (U.S.); National Center for HIV, STD, and TB Prevention (U.S.), Division of AIDS, STD, and TB Laboratory Research.; National Center on Birth Defects and Developmental Disabilities (Centers for Disease Control and Prevention);

Published:

January 14, 2004

Series:

MMWR. Recommendations and reports : Morbidity and mortality weekly report. Recommendations and reports ; v. 53, no. RR-1

Description:

Primary immunodeficiency (PI) diseases are a group of primarily single-gene disorders of the immune system. Approximately 100 separate PI diseases have been described, but <20 probably account for >90% of cases. Although diverse, PI diseases share th...

File Type:

[PDF - 728.41 KB]
Whole-brain activity mapping onto a zebrafish brain atlas

Personal Author:

Randlett, Owen; Wee, Caroline L.; Naumann, Eva A.; Nnaemeka, Onyeka; Schoppik, David; Fitzgerald, James E.; Portugues, Ruben; Lacoste, Alix M.B.; Riegler, Clemens; Engert, Florian; Schier, Alexander F.;

Published:

Nov 2015

Source:

Nat Methods. 12(11):1039-1046.

Description:

In order to localize the neural circuits involved in generating behaviors, it is necessary to assign activity onto anatomical maps of the nervous system. Using brain registration across hundreds of larval zebrafish, we have built an expandable open-s...

File Type:

[PDF - 2.52 MB]