A Proteomics Approach to Profiling the Temporal Translational Response to Stress and Growth
Name
1-s2.0-S2589004218301949-main.pdf
Description
Published version
Size
7.15 MB
Format
Adobe PDF
Checksum (MD5)
60cfd9d8a101083c04abc4c707206532
Author(s)
Rothenberg, Daniel Abram
Taliaferro, Jefferson Matthew
Huber, Sabrina M
Dedon, Peter C
White, Forest M.
Date Issued
November 2018
Journal
iScience
Publisher
Elsevier BV
Citation
Rothenberg, Daniel A. et al. “A Proteomics Approach to Profiling the Temporal Translational Response to Stress and Growth.” iScience, vol. 9, 2018, pp. 367-381 © 2018 The Author(s)
Version
Final published version
Abstract
To quantify dynamic protein synthesis rates, we developed MITNCAT, a method combining multiplexed isobaric mass tagging with pulsed SILAC (pSILAC)and bio-orthogonal non-canonical amino acid tagging (BONCAT)to label newly synthesized proteins with azidohomoalanine (Aha), thus enabling high temporal resolution across multiple conditions in a single analysis. MITNCAT quantification of protein synthesis rates following induction of the unfolded protein response revealed global down-regulation of protein synthesis, with stronger down-regulation of glycolytic and protein synthesis machinery proteins, but up-regulation of several key chaperones. Waves of temporally distinct protein synthesis were observed in response to epidermal growth factor, with altered synthesis detectable in the first 15 min. Comparison of protein synthesis with mRNA sequencing and ribosome footprinting distinguished protein synthesis driven by increased transcription versus increased translational efficiency. Temporal delays between ribosome occupancy and protein synthesis were observed and found to correlate with altered codon usage in significantly delayed proteins.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Biology
Koch Institute for Integrative Cancer Research at MIT
Terms of Use
Creative Commons Attribution 4.0 International license
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1016/J.ISCI.2018.11.004
