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Matteo Perino, Per Haberkant, Frank Stein, Raquel Marco-Ferreres, Kathleen-Una Towns, Mikhail Savitski, Eileen E. M. Furlong (EMBL, Heidelberg, Germany)
During the initial phases of embryonic development, the zygotic genome is kept silent and embryonic development is exclusively sustained by maternally provided RNA and proteins. Only later the embryo will start transcribing its own RNA during Zygotic Genome Activation (ZGA). All the factors required to activate the silent zygotic genome must, by definition, be maternally provided, thus present from fertilization. Yet ZGA happens many hours and cell cycles later, and with remarkable precision. Thermal Proteome Profiling (TPP), a technique that estimate changes in protein activity by measuring thermal stability provides an unexplored avenue to probe ZGA regulatory mechanisms, by going beyond simple protein quantification. We optimized TPP for live, developing embryo and found that 7.2% of the embryonic proteome shows stability changes before or upon ZGA, including Zelda, the most well-studied player in Drosophila ZGA. We shortlisted both known and uncharacterized genes, and we are characterising their function by germline-specific genetic depletion.https://www.embl.de/research/units/genome_biology/furlong/index.html