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.

Public Health Relevance

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.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
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Special Emphasis Panel (ZRG1-GGG-A (50))
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Hamlet, Michelle R
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Harvard University
Schools of Medicine
United States
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Boke, Elvan; Ruer, Martine; Wühr, Martin et al. (2016) Amyloid-like Self-Assembly of a Cellular Compartment. Cell 166:637-50
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
Klein, Allon M; Mazutis, Linas; Akartuna, Ilke et al. (2015) Droplet barcoding for single-cell transcriptomics applied to embryonic stem cells. Cell 161:1187-201
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
Erickson, Brian K; Jedrychowski, Mark P; McAlister, Graeme C et al. (2015) Evaluating multiplexed quantitative phosphopeptide analysis on a hybrid quadrupole mass filter/linear ion trap/orbitrap mass spectrometer. Anal Chem 87:1241-9
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
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
Wühr, Martin; Güttler, Thomas; Peshkin, Leonid et al. (2015) The Nuclear Proteome of a Vertebrate. Curr Biol 25:2663-71
Gujral, Taranjit S; Chan, Marina; Peshkin, Leonid et al. (2014) A noncanonical Frizzled2 pathway regulates epithelial-mesenchymal transition and metastasis. Cell 159:844-56
Ishihara, Keisuke; Nguyen, Phuong A; Wühr, Martin et al. (2014) Organization of early frog embryos by chemical waves emanating from centrosomes. Philos Trans R Soc Lond B Biol Sci 369:

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