Expression patterns of CREB binding protein (CREBBP) and its methylated species during zebrafish development

Abstract

Proper embryonic development requires a fine-tuned control of gene expression, which is achieved in part through the activity of transcription coactivators or corepressors. The nuclear coactivator cAMP-response element-binding protein (CREB) binding protein (CREBBP or CBP) interacts with numerous transcription factors and thereby plays a key role in various signaling pathways. Interestingly, in cell-based studies CREBBP activity is modulated by post-translational modifications such as methylation on arginine residues which is catalyzed by coactivator-associated arginine methyltransferase 1 (CARM1). However, whether and where CREBBP, and in particular its methylated forms, are expressed during development in vertebrates has not been addressed so far. Here, we analyzed the expression of the two crebbp genes (crebbpa & crebbpb) during zebrafish development using both RT-qPCR and in situ hybridization. We found that while crebbpa expression is higher in posterior, caudal nascent somites during somitogenesis, crebbpb accumulates in anterior, rostral, and more mature somites. In addition, crebbpa mRNA is enriched in the central myotome at 24 hpf indicating that its expression is spatially and temporally controlled. We next characterized the expression of CREBBP protein from blastula to gastrula stages by immunohistochemistry. We found that while CREBBP is clearly cytoplasmic in the early blastula, it becomes both cytoplasmic and nuclear at 30% epiboly before turning mainly nuclear during gastrulation. Of interest, CREBBP methylated species appear to be mainly nuclear from 30% epiboly to 6-somite stage. This suggests that methylation may regulate CREBBP import to the nucleus during zebrafish development and could therefore participate in the control of early developmental processes.

Widespread Enhancer Dememorization and Promoter Priming during Parental-to-Zygotic Transition

“…And the doses of HATs-MOs were 4 ng per oocyte for each MO, and dose of cMO in this assay was 12 ng per oocyte. The sequences of these MOs were 5 0 -ACAATGAGGTCTTGGTAGGCATTTC-3 0 (dnmt1-MO) (Rai et al, 2006), 5 0 -GACTGTTGCCACCTGCCATGCCCAT À3 0 (crebbpa-MO) (Batut et al, 2015), 5 0 -TCCAGTAGATTATCAGCCATCGTGC À3 0 (crebbpb-MO), 5 0 -GGAGTCCAGCACATTATCGGC CATA À3 0 (ep300b-MO), and 5 0 -CCTCTTACCTCAGTTACAATTTATA-3 0 (cMO). MOs were dissolved in RNase-free water, and heated to 65 C for 10 min before microinjection.…”

Epigenetic Regulators Modulate Muscle Damage in Duchenne Muscular Dystrophy Model

“…CBP was shown to functionally activate MyoD by acetylation and to directly interact with MEF2C (Giordani and Puri, 2013). CBP expression in zebrafish muscle was reported recently (Batut et al, 2015). We used the zebrafish model, extensively characterised and widely used for studying muscular dystrophy (Bajanca et al, 2015), to explore the effects of overexpressing CBP in a Dystrophin-null background.…”

Μελέτη του λειτουργικού ρόλου της πρωτεΐνης Set κατά τα πρώιμα αναπτυξιακά στάδια του οργανισμού-μοντέλου Danio rerio (zebrafish)

“…Στο zebrafish υπάρχουν δύο γονίδια crebbp (crebbpa και crebbpb) που κωδικοποιούν τις ορθόλογες πρωτεΐνες της CBP. Η ανάλυση του προτύπου έκφρασης των mRNA των δύο γονιδίων κατά την εμβρυογένεση, αποκάλυψε έντονη διάχυτη κατανομή η οποία μέχρι τις 24 hpf είχε περιοριστεί στον οφθαλμό και το ωτικό κυστίδιο (Batut et al, 2015), υποδηλώνοντας βασικό ρόλο των συγκεκριμένων γονιδίων στην διαφοροποίηση αυτών των οργάνων. Το παρόν εύρημα συνδυαστικά αφενός με το αντίστοιχο πρότυπο έκφρασης των seta/b και αφετέρου με τη σχέση αλληλεπίδρασης μεταξύ SET και CBP, ενισχύει την υπόθεση της δράσης των υπό μελέτη γονιδίων στη μεταγραφική ρύθμιση παραγόντων που σχετίζονται με τη νευρική διαφοροποίηση, μέσω των CBP/p300.…”