Article thumbnail

A Proteomics Approach to Profiling the Temporal Translational Response to Stress and Growth

By Daniel A. Rothenberg, J. Matthew Taliaferro, Sabrina M. Huber, Thomas J. Begley, Peter C. Dedon and Forest M. White

Abstract

Summary: 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. : Functional Aspects of Cell Biology; Methodology in Biological Sciences; Proteomics Subject Areas: Functional Aspects of Cell Biology, Methodology in Biological Sciences, Proteomic

Topics: Science, Q
Publisher: Elsevier
Year: 2018
DOI identifier: 10.1016/j.isci.2018.11.004
OAI identifier: oai:doaj.org/article:944a04c5a2954b2c94e120526e239a86
Journal:
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • https://doaj.org/toc/2589-0042 (external link)
  • http://www.sciencedirect.com/s... (external link)
  • https://doaj.org/article/944a0... (external link)
  • Suggested articles


    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.