UHRF1 regulation of Dnmt1 is required for pre-gastrula zebrafish development

Brandon Kent, Elena Magnani, Martin J. Walsh, Kirsten Sadler Edepli

    Research output: Contribution to journalArticle

    Abstract

    Landmark epigenetic events underlie early embryonic development, yet how epigenetic modifiers are regulated to achieve rapid epigenome re-patterning is not known. Uhrf1 and DNA methyltransferase 1 (Dnmt1) are known to largely mediate maintenance DNA methylation and Uhrf1 is also required for both Dnmt1 localization and stability. Here, we investigate how these two key epigenetic modifiers regulate early zebrafish development and characterize the developmental consequences of disrupting their homeostatic relationship. Unlike Uhrf1 knockdown, which causes developmental arrest and death prior to gastrulation, overexpression of human UHRF1 (WT-UHRF1) caused asymmetric epiboly, inefficient gastrulation and multi-systemic defects. UHRF1 phosphorylation was previously demonstrated as essential for zebrafish embryogenesis, and we found that penetrance of the asymmetric epiboly phenotype was significantly increased in embryos injected with mRNA encoding non-phosphorylatable UHRF1 (UHRF1S661A). Surprisingly, both WT-UHRF1 and UHRF1S661A overexpression caused DNA hypomethylation. However, since other approaches that caused an equivalent degree of DNA hypomethylation did not cause the asymmetric epiboly phenotype, we conclude that bulk DNA methylation is not the primary mechanism. Instead, UHRF1S661A overexpression resulted in accumulation of Dnmt1 protein and the overexpression of both WT and a catalytically inactive Dnmt1 phenocopied the assymetric epiboly phenotype. Dnmt1 knockdown suppressed the phenotype caused by UHRF1S661A overexpression, and Uhrf1 knockdown suppressed the effect of Dnmt1 overexpression. Therefore, we conclude that the interaction between these two proteins is the mechanism underlying the gastrulation defects. This indicates that Dnmt1 stability requires UHRF1 phosphorylation and that crosstalk between the proteins is essential for the function of these two important epigenetic regulators during gastrulation.

    Original languageEnglish (US)
    Pages (from-to)99-113
    Number of pages15
    JournalDevelopmental Biology
    Volume412
    Issue number1
    DOIs
    StatePublished - Apr 1 2016

    Fingerprint

    Gastrula
    Methyltransferases
    Zebrafish
    DNA
    Gastrulation
    Epigenomics
    Phenotype
    DNA Methylation
    Embryonic Development
    Protein Methyltransferases
    Phosphorylation
    Penetrance
    Proteins
    Embryonic Structures
    Maintenance
    Messenger RNA

    Keywords

    • DNA methylation
    • Dnmt1
    • Epiboly
    • Epigenetics
    • Uhrf1
    • Zebrafish development

    ASJC Scopus subject areas

    • Molecular Biology
    • Developmental Biology
    • Cell Biology

    Cite this

    UHRF1 regulation of Dnmt1 is required for pre-gastrula zebrafish development. / Kent, Brandon; Magnani, Elena; Walsh, Martin J.; Sadler Edepli, Kirsten.

    In: Developmental Biology, Vol. 412, No. 1, 01.04.2016, p. 99-113.

    Research output: Contribution to journalArticle

    Kent, Brandon ; Magnani, Elena ; Walsh, Martin J. ; Sadler Edepli, Kirsten. / UHRF1 regulation of Dnmt1 is required for pre-gastrula zebrafish development. In: Developmental Biology. 2016 ; Vol. 412, No. 1. pp. 99-113.
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