[en] ARP/ASCL transcription factors are key determinants of cell fate specification in a wide variety of tissues, coordinating the acquisition of generic cell fates and of specific subtype identities. How these factors, recognizing highly similar DNA motifs, display specific activities, is not yet fully understood. To address this issue, we overexpressed different ARP/ASCL factors in zebrafish ascl1a-/- mutant embryos to determine which ones are able to rescue the intestinal secretory lineage. We found that Ascl1a/b, Atoh1a/b and Neurod1 factors are all able to trigger the first step of the secretory regulatory cascade but distinct secretory cells are induced by these factors. Indeed, Neurod1 rescues the enteroendocrine lineage while Ascl1a/b and Atoh1a/b rescue the goblet cells. Gain-of-function experiments with Ascl1a/Neurod1 chimeric proteins revealed that the functional divergence is encoded by a 19-aa ultra-conserved element (UCE), present in all Neurod members but absent in the other ARP/ASCL proteins. Importantly, inserting the UCE into the Ascl1a protein reverse the rescuing capacity of this Ascl1a chimeric protein that cannot rescue the goblet cells anymore but efficiently rescue the enteroendocrine cells. This novel domain acts indeed as a goblet cell fate repressor that inhibits gfi1aa expression, known to be important for goblet cell differentiation. Deleting the UCE domain of the endogenous Neurod1 protein leads to an increase in the number of goblet cells concomitant with a reduction of the enteroendocrine cells, phenotype also observed in the neurod1 null mutant. This highlight the crucial function of the UCE domain for NeuroD1 activity in the intestine. As Gfi1 acts as a binary cell fate switch in several tissues where Neurod1 is also expressed, we can envision a similar role of the UCE in other tissues, allowing Neurod1 to repress Gfi1 to influence the balance between cell fates.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Reuter, Anne-Sophie ; Université de Liège - ULiège > GIGA > GIGA Stem Cells - Zebrafish Development and Disease Model
Stern, David ; Université de Liège - ULiège > GIGA > GIGA Platform Genomics ; Laboratory of Zebrafish Development and Disease Models (ZDDM), GIGA, University of Liège - ULiège
Bernard, Alice ; Université de Liège - ULiège > Integrative Biological Sciences (InBioS) ; Laboratory of Zebrafish Development and Disease Models (ZDDM), GIGA, University of
Goossens, Chiara ; Université de Liège - ULiège > Département des sciences de la vie ; Laboratory of Zebrafish Development and Disease Models (ZDDM), GIGA, University of Liège - ULiège
Lavergne, Arnaud ; Université de Liège - ULiège > Département de gestion vétérinaire des Ressources Animales (DRA) > Génomique animale ; Laboratory of Zebrafish Development and Disease Models (ZDDM), GIGA, University of Liège - ULiège
Flasse, Lydie ; Université de Liège - ULiège > GIGA > GIGA Stem Cells - Zebrafish Development and Disease Model ; Laboratory of Zebrafish Development and Disease Models (ZDDM), GIGA, University of
Von Berg, Virginie ; Centre Hospitalier Universitaire de Liège - CHU > > Service de génétique ; Laboratory of Zebrafish Development and Disease Models (ZDDM), GIGA, University of
Manfroid, Isabelle ; Université de Liège - ULiège > GIGA > GIGA Stem Cells - Zebrafish Development and Disease Model ; Laboratory of Zebrafish Development and Disease Models (ZDDM), GIGA, University of Liège - ULiège
Peers, Bernard ; Université de Liège - ULiège > GIGA > GIGA Stem Cells - Zebrafish Development and Disease Model ; Laboratory of Zebrafish Development and Disease Models (ZDDM), GIGA, University of Liège - ULiège
Voz, Marianne ; Université de Liège - ULiège > GIGA > GIGA Stem Cells - Zebrafish Development and Disease Model ; Laboratory of Zebrafish Development and Disease Models (ZDDM), GIGA, University of Liège - ULiège
Language :
English
Title :
Identification of an evolutionarily conserved domain in Neurod1 favouring enteroendocrine versus goblet cell fate.
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