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Conserved enhancer logic controls the notochord expression of vertebrate Brachyury

Kemmler, C. L.; Smolikova, J.; Moran, H. R.; Mannion, B. J.; Knapp, D.; Lim, F.; Czarkwiani, A.; Hermosilla Aguayo, V.; Rapp, V.; Fitch, O. E.; Boetschi, S.; Selleri, L.; Farley, E.; Braasch, I.; Yun, M.; Visel, A.; Osterwalder, M.; Mosimann, C.; Kozmik, Z.; Burger, A.

2023-04-20 genetics
10.1101/2023.04.20.536761 bioRxiv
Show abstract

The cell type-specific expression of key transcription factors is central to development. Brachyury/T/TBXT is a major transcription factor for gastrulation, tailbud patterning, and notochord formation; however, how its expression is controlled in the mammalian notochord has remained elusive. Here, we identify the complement of notochord-specific enhancers in the mammalian Brachyury/T/TBXT gene. Using transgenic assays in zebrafish, axolotl, and mouse, we discover three Brachyury-controlling notochord enhancers T3, C, and I in human, mouse, and marsupial genomes. Acting as Brachyury-responsive, auto-regulatory shadow enhancers, deletion of all three enhancers in mouse abolishes Brachyury/T expression selectively in the notochord, causing specific trunk and neural tube defects without gastrulation or tailbud defects. Sequence and functional conservation of Brachyury-driving notochord enhancers with the brachyury/tbxtb loci from diverse lineages of fishes dates their origin to the last common ancestor of jawed vertebrates. Our data define the enhancers for Brachyury/T/TBXTB notochord expression as ancient mechanism in axis development.

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