Proteomics has special significance for Xenopus research because key events in meiosis, fertilization and early development are regulated at the level of translation, post- translational modification and proteolysis, and not by transcription. Proteomics is the only tool for systematically investigating these levels of regulation and discovering new mechanisms. We propose to develop a cutting edge proteomic research platform for Xenopus embryos and egg extract by migrating technology developed in human and yeast systems, and to publicly disseminate the databases and tools we develop. We will also optimize methods for proteomic analysis of protein complexes and compartments in egg extract. Using these tools we will quantify relative amounts of >5,000 proteins, >10,000 phosphorylation sites and >1,500 ubiquitination sites in early development and in egg extracts under normal and perturbed conditions. We will investigate the mechanism of cell cycle regulation, how the cell cycle changes during early development, how microtubules are nucleated, and how embryos detect changes in the nucleus to cytoplasm ratio.
Project Narrative We will develop broad based tools for the study of changes in the protein levels during the cell division cycle, bringing the revolutionary technique of mass spectrometry to embryonic systems. Such studies could offer a better basis for designing drugs that affect important pathways in cancer and inflammation.
| Gujral, Taranjit S; Kirschner, Marc W (2017) Hippo pathway mediates resistance to cytotoxic drugs. Proc Natl Acad Sci U S A 114:E3729-E3738 |
| Erickson, Brian K; Rose, Christopher M; Braun, Craig R et al. (2017) A Strategy to Combine Sample Multiplexing with Targeted Proteomics Assays for High-Throughput Protein Signature Characterization. Mol Cell 65:361-370 |
| Presler, Marc; Van Itallie, Elizabeth; Klein, Allon M et al. (2017) Proteomics of phosphorylation and protein dynamics during fertilization and meiotic exit in the Xenopus egg. Proc Natl Acad Sci U S A 114:E10838-E10847 |
| Hanschen, Erik R; Marriage, Tara N; Ferris, Patrick J et al. (2016) The Gonium pectorale genome demonstrates co-option of cell cycle regulation during the evolution of multicellularity. Nat Commun 7:11370 |
| Boke, Elvan; Ruer, Martine; Wühr, Martin et al. (2016) Amyloid-like Self-Assembly of a Cellular Compartment. Cell 166:637-650 |
| Mitchison, Timothy J; Ishihara, Keisuke; Nguyen, Phuong et al. (2015) Size Scaling of Microtubule Assemblies in Early Xenopus Embryos. Cold Spring Harb Perspect Biol 7:a019182 |
| Wühr, Martin; Güttler, Thomas; Peshkin, Leonid et al. (2015) The Nuclear Proteome of a Vertebrate. Curr Biol 25:2663-71 |
| Peshkin, Leonid; Wühr, Martin; Pearl, Esther et al. (2015) On the Relationship of Protein and mRNA Dynamics in Vertebrate Embryonic Development. Dev Cell 35:383-94 |
| Braun, Craig R; Bird, Gregory H; Wühr, Martin et al. (2015) Generation of multiple reporter ions from a single isobaric reagent increases multiplexing capacity for quantitative proteomics. Anal Chem 87:9855-63 |
| Klein, Allon M; Mazutis, Linas; Akartuna, Ilke et al. (2015) Droplet barcoding for single-cell transcriptomics applied to embryonic stem cells. Cell 161:1187-1201 |
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