NDLI: Use of conditioned media is critical for studies of regulation in response to rapid heat shock

Content Provider	Springer Nature Link
Author	Mahat, Dig B. Lis, John T.
Copyright Year	2016
Abstract	Heat shock response (HSR) maintains and restores protein homeostasis when cells are exposed to proteotoxic heat stress. Heat shock (HS) triggers a rapid and robust change in genome-wide transcription, protein synthesis, and chaperone activity; and therefore, the HSR has been widely used as a model system in these studies. The conventional method of performing instantaneous HS in the laboratory uses heated fresh media to induce HSR when added to cells. However, addition of fresh media to cells may evoke additional cellular responses and signaling pathways. Here, we compared the change in global transcription profile when HS is performed with either heated fresh media or heated conditioned media. We found that the use of heated fresh media induces transcription of hundreds of genes that HS alone does not induce, and masks or partially masks HS-mediated downregulation of thousands of genes. The fresh-media-dependent upregulated genes encode ribosomal subunit proteins involved in translation and RNA processing factors. More importantly, fresh media also induce transcription of several heat shock protein genes (Hsps) in a heat shock factor 1 (HSF1)-independent manner. Thus, we conclude that a conventional method of HS with heated fresh media causes changes in transcription regulation that confound the actual change caused solely by elevated temperature of cells.
Starting Page	155
Ending Page	162
Page Count	8
File Format	PDF
ISSN	13558145
Journal	Cell Stress and Chaperones
Volume Number	22
Issue Number	1
e-ISSN	14661268
Language	English
Publisher	Springer Netherlands
Publisher Date	2016-11-03
Publisher Place	Dordrecht
Access Restriction	Subscribed
Subject Keyword	Heat shock Heat shock proteins Transcription Conditioned media Gene regulation PRO-seq Serum Glucose Cell culture media Biomedicine Cell Biology Biochemistry Immunology Cancer Research Neurosciences
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry

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