Combining Genetic Perturbations and Proteomics to Examine Kinase-Phosphatase Networks in Drosophila Embryos
Author
Sopko, Richelle
Foos, Marianna
Vinayagam, Arunachalam
Zhai, Bo
Randklev, Sakara
Perkins, Lizabeth A.
Published Version
https://doi.org/10.1016/j.devcel.2014.07.027Metadata
Show full item recordCitation
Sopko, Richelle, Marianna Foos, Arunachalam Vinayagam, Bo Zhai, Richard Binari, Yanhui Hu, Sakara Randklev et al. "Combining Genetic Perturbations and Proteomics to Examine Kinase-Phosphatase Networks in Drosophila Embryos." Developmental Cell 31, no. 1 (2014): 114-127. DOI: 10.1016/j.devcel.2014.07.027Abstract
Connecting phosphorylation events to kinases and phosphatases is key to understanding the molecular organization and signaling dynamics of networks. We have generated a validated set of transgenic RNA-interference reagents for knockdown and characterization of all protein kinases and phosphatases present during early Drosophila melanogaster development. These genetic tools enable collection of sufficient quantities of embryos depleted of single gene products for proteomics. As a demonstration of an application of the collection, we have used multiplexed isobaric-labeling for quantitative proteomics to derive global phosphorylation signatures associated with kinase-depleted embryos, in order to systematically link phosphosites with relevant kinases. We demonstrate how this strategy uncovers kinase consensus motifs and prioritizes phosphoproteins for kinase target validation. We validate this approach by providing auxiliary evidence for Wee kinase-directed regulation of the chromatin regulator Stonewall. Further, we show how correlative phosphorylation at the site level can indicate function, as exemplified by Sterile20-like kinase-dependent regulation of Stat92E.Other Sources
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4208667/Terms of Use
This article is made available under the terms and conditions applicable to Open Access Policy Articles, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#OAPCitable link to this page
https://nrs.harvard.edu/URN-3:HUL.INSTREPOS:37368029
Collections
- HMS Scholarly Articles [17922]
Contact administrator regarding this item (to report mistakes or request changes)