1. Krox20 hindbrain regulation incorporates multiple modes of cooperation between cis-acting elements
- Author
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Elodie, Thierion, Johan, Le Men, Samuel, Collombet, Céline, Hernandez, Fanny, Coulpier, Patrick, Torbey, Morgane, Thomas-Chollier, Daan, Noordermeer, Patrick, Charnay, Pascale, Gilardi-Hebenstreit, Institut de biologie de l'ENS Paris (IBENS), Département de Biologie - ENS Paris, École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Sorbonne Université - Institut de Formation Doctorale (IFD ), Sorbonne Université (SU), Institut de Biologie Intégrative de la Cellule (I2BC), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), The P.C. laboratory was financed by the Institut National de la Recherche Médicale, the Centre National de la Recherche Scientifique, the Ministère de la Recherche et Technologie and the Fondation pour la Recherche Médicale (FRM). It has received support under the PIA launched by the French Government and implemented by the ANR, with the references: ANR-10-LABX-54 MEMOLIFE and ANR-11-IDEX-0001-02 PSL Research University. The genomic platform of the IBENS is supported by France Génomique national infrastructure and funded as part of the Programme 'Investissements d'Avenir' (PIA) managed by the Agence Nationale de la Recherche (ANR, contract ANR-10-INBS-0009). ET and JLM were supported by doctoral grants from Sorbonne Universités and by the FRM. PT is supported by doctoral grant from PSL Research University., ANR-10-IDEX-0001,PSL,Paris Sciences et Lettres(2010), ANR-11-IDEX-0001,Amidex,INITIATIVE D'EXCELLENCE AIX MARSEILLE UNIVERSITE(2011), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), Institut de biologie de l'ENS Paris (UMR 8197/1024) (IBENS), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Biologie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Bodescot, Myriam, Initiative d'excellence - Paris Sciences et Lettres - - PSL2010 - ANR-10-IDEX-0001 - IDEX - VALID, INITIATIVE D'EXCELLENCE AIX MARSEILLE UNIVERSITE - - Amidex2011 - ANR-11-IDEX-0001 - IDEX - VALID, and Organisation et montée en puissance d'une Infrastructure Nationale de Génomique - - France-Génomique2010 - ANR-10-INBS-0009 - INBS - VALID
- Subjects
Embryology ,Enhancer Elements ,lcsh:QH426-470 ,DNA transcription ,Gene Expression ,Sequence Homology, Nucleic Acid ,[SDV.BDD] Life Sciences [q-bio]/Development Biology ,Genetics ,[SDV.BBM] Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Animals ,Gene Regulation ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Regulatory Elements, Transcriptional ,[SDV.BDD]Life Sciences [q-bio]/Development Biology ,Body Patterning ,Early Growth Response Protein 1 ,Mice, Knockout ,Mammalian Genomics ,Chromosome Biology ,Embryos ,Gene Expression Regulation, Developmental ,Biology and Life Sciences ,Cell Biology ,Genomics ,Chromatin ,Rhombencephalon ,lcsh:Genetics ,Enhancer Elements, Genetic ,Animal Genomics ,Genetic Loci ,Mutation ,Epigenetics ,Research Article ,Developmental Biology - Abstract
Developmental genes can harbour multiple transcriptional enhancers that act simultaneously or in succession to achieve robust and precise spatiotemporal expression. However, the mechanisms underlying cooperation between cis-acting elements are poorly documented, notably in vertebrates. The mouse gene Krox20 encodes a transcription factor required for the specification of two segments (rhombomeres) of the developing hindbrain. In rhombomere 3, Krox20 is subject to direct positive feedback governed by an autoregulatory enhancer, element A. In contrast, a second enhancer, element C, distant by 70 kb, is active from the initiation of transcription independent of the presence of the KROX20 protein. Here, using both enhancer knock-outs and investigations of chromatin organisation, we show that element C possesses a dual activity: besides its classical enhancer function, it is also permanently required in cis to potentiate the autoregulatory activity of element A, by increasing its chromatin accessibility. This work uncovers a novel, asymmetrical, long-range mode of cooperation between cis-acting elements that might be essential to avoid promiscuous activation of positive autoregulatory elements., Author summary The formation of multicellular organisms from the egg to the adult stage is largely under genetic control. The activation of specific genes is governed by regulatory DNA sequences present nearby on the chromosome. Most of these sequences promote activation and are called enhancers. In this paper, we study two enhancers governing the expression of a gene involved in the formation of the posterior brain in vertebrates. One of these enhancers is involved in a positive feedback loop: it is itself activated by the protein product of the gene that it regulates. The other enhancer was thought to be only involved in the initial accumulation of the protein, necessary for the subsequent activation of the feedback loop. Here we show that the second enhancer directly cooperates with the autoregulatory enhancer to increase its accessibility and its activity. Our work uncovers a novel, long-range mode of cooperation between enhancers that restricts the domain of action of autoregulatory enhancers within embryos and might be essential to avoid their inappropriate activation.
- Published
- 2017
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