Regensburg 2016 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 18: Posters - DNA, RNA and Related Enzymes
BP 18.4: Poster
Montag, 7. März 2016, 17:30–19:30, Poster C
Subnuclear Microarchitecture is Established when Transcription is Activated in Zebrafish Embryos — •Lennart Hilbert1,2,3, Yuko Sato4, Hiroshi Kimura4, Alf Honigmann3, Vasily Zaburdaev2, and Nadine Vastenhouw3 — 1Center for Systems Biology Dresden — 2MPI for Physics of Complex Systems — 3MPI of Molecular Cell Biology and Genetics — 4Tokyo Institute of Technology
DNA transcription is a fundamental process of cellular function. Still, the driving forces of spatial organization of the transcription machinery in the nucleus remain poorly understood. Here, we used the onset of transcription in zebrafish embryos as a model system to investigate the contribution of transcription to spatial organization. To enable super-resolution microscopy of subnuclear organization, we dissociated embryos into individual cells. Clones of these cells exhibited transcription onset as seen in embryos. We imaged DNA and active RNA polymerase II (Pol II) in fixed clones by widefield and 3D STED super-resolution microscopy. DNA was first homogeneously distributed but segregated into spatially confined domains after transcription onset. Pol II foci with a granular sub-micron structure were seen. Focus frequency and structural complexity increased with intensifying transcription. After transcription onset, a nucleus-wide, interconnected network of Pol II-compartments formed. Live cell Pol II detection with fluorescence-tagged antibody fragments reproduced the fixed cell results. The techniques enabled by embryo dissociation will support comprehensive assessment of transcription as a driver of subnuclear organization.