Title | A Proteomics Approach to Profiling the Temporal Translational Response to Stress and Growth. |
Publication Type | Journal Article |
Year of Publication | 2018 |
Authors | Rothenberg DA, J Taliaferro M, Huber SM, Begley TJ, Dedon PC, White FM |
Journal | iScience |
Volume | 9 |
Pagination | 367-381 |
Date Published | 2018 Nov 30 |
ISSN | 2589-0042 |
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. |
DOI | 10.1016/j.isci.2018.11.004 |
Alternate Journal | iScience |
PubMed ID | 30466063 |
PubMed Central ID | PMC6249402 |
Grant List | P42 ES027707 / ES / NIEHS NIH HHS / United States R01 ES026856 / ES / NIEHS NIH HHS / United States P30 CA014051 / CA / NCI NIH HHS / United States R01 ES024615 / ES / NIEHS NIH HHS / United States P30 ES002109 / ES / NIEHS NIH HHS / United States P01 CA026731 / CA / NCI NIH HHS / United States R01 CA096504 / CA / NCI NIH HHS / United States U54 CA210180 / CA / NCI NIH HHS / United States |