NDLI: Absolute quantification of transcription factors during cellular differentiation using multiplexed targeted proteomics.

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Absolute quantification of transcription factors during cellular differentiation using multiplexed targeted proteomics.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Simicevic, Jovan Schmid, Adrien W. Gilardoni, Paola A. Zoller, Benjamin Raghav, Sunil K. Krier, Irina Gubelmann, Carine Lisacek, Frédérique Naef, Felix Moniatte, Marc Deplancke, Bart
Description	Country affiliation: Switzerland Author Affiliation: Simicevic J ( Laboratory of Systems Biology and Genetics, Institute of Bioengineering, School of Life Sciences, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.)
Abstract	The cellular abundance of transcription factors (TFs) is an important determinant of their regulatory activities. Deriving TF copy numbers is therefore crucial to understanding how these proteins control gene expression. We describe a sensitive selected reaction monitoring-based mass spectrometry assay that allowed us to determine the copy numbers of up to ten proteins simultaneously. We applied this approach to profile the absolute levels of key TFs, including PPARÎ³ and RXR , during terminal differentiation of mouse 3T3-L1 pre-adipocytes. Our analyses revealed that individual TF abundance differs dramatically (from â¼250 to >300,000 copies per nucleus) and that their dynamic range during differentiation can vary up to fivefold. We also formulated a DNA binding model for PPARÎ³ based on TF copy number, binding energetics and local chromatin state. This model explains the increase in PPARÎ³ binding sites during the final differentiation stage that occurs despite a concurrent saturation in PPARÎ³ copy number.
File Format	HTM / HTML
ISSN	15487091
Issue Number	6
Volume Number	10
e-ISSN	15487105
Journal	Nature Methods
Language	English
Publisher	Nature Publishing Group
Publisher Date	2013-06-01
Publisher Place	United States
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Discipline Clinical Laboratory Techniques Cell Differentiation Proteomics Methods Transcription Factors Analysis 3t3-l1 Cells Animals Dna Metabolism Mice Ppar Gamma Retinoid X Receptor Alpha Journal Article Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Molecular Biology Biotechnology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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Absolute quantification of transcription factors during cellular differentiation using multiplexed targeted proteomics.

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Absolute quantification of transcription factors during cellular differentiation using multiplexed targeted proteomics.