UHRF1 phosphorylation by cyclin A2/cyclin-dependent kinase 2 is required for zebrafish embryogenesis

Jaime Chua, Elizabeth A. Loughlin, Naseem A. Gaur, Sucharita SenBanerjee, Vinitha Jacob, Christopher Monson, Brandon Kent, Amanke Oranu, Yuanying Ding, Chinweike Ukomadu, Kirsten Sadler Edepli

Research output: Contribution to journalArticle

Abstract

Ubiquitin-like, containing PHD and RING finger domains 1 (uhrf1) is regulated at the transcriptional level during the cell cycle and in developing zebrafish embryos. We identify phosphorylation as a novel means of regulating UHRF1 and demonstrate that Uhrf1 phosphorylation is required for gastrulation in zebrafish. Human UHRF1 contains a conserved cyclin-dependent kinase 2 (CDK2) phosphorylation site at Ser-661 that is phosphorylated in vitro by CDK2 partnered with cyclin A2 (CCNA2), but not cyclin E. An antibody specific for phospho-Ser-661 recognizes UHRF1 in both mammalian cancer cells and in nontransformed zebrafish cells, but not in zebrafish bearing a mutation in ccna2. Depleting Uhrf1 from zebrafish embryos by morpholino injection causes arrest before gastrulation and early embryonic death. This phenotype is rescued by wild-type UHRF1, but not by UHRF1 in which the phospho-acceptor site is mutated, demonstrating that UHRF1 phosphorylation is essential for embryogenesis. UHRF1 was detected in the nucleus and cytoplasm, whereas nonphosphorylatable UHRF1 is unable to localize to the cytoplasm, suggesting the importance of localization in UHRF1 function. Together, these data point to an essential role for UHRF1 phosphorylation by CDK/CCNA2 during early vertebrate development.

Original languageEnglish (US)
Pages (from-to)59-70
Number of pages12
JournalMolecular Biology of the Cell
Volume23
Issue number1
DOIs
StatePublished - Jan 1 2012

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Cyclin A2
Cyclin-Dependent Kinase 2
Zebrafish
Embryonic Development
Phosphorylation
Gastrulation
Cytoplasm
Embryonic Structures
RING Finger Domains
Phospho-Specific Antibodies
Morpholinos
Cyclin E
Ubiquitin
Vertebrates
Cell Cycle
Phenotype
Mutation
Injections
Neoplasms

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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UHRF1 phosphorylation by cyclin A2/cyclin-dependent kinase 2 is required for zebrafish embryogenesis. / Chua, Jaime; Loughlin, Elizabeth A.; Gaur, Naseem A.; SenBanerjee, Sucharita; Jacob, Vinitha; Monson, Christopher; Kent, Brandon; Oranu, Amanke; Ding, Yuanying; Ukomadu, Chinweike; Sadler Edepli, Kirsten.

In: Molecular Biology of the Cell, Vol. 23, No. 1, 01.01.2012, p. 59-70.

Research output: Contribution to journalArticle

Chua, J, Loughlin, EA, Gaur, NA, SenBanerjee, S, Jacob, V, Monson, C, Kent, B, Oranu, A, Ding, Y, Ukomadu, C & Sadler Edepli, K 2012, 'UHRF1 phosphorylation by cyclin A2/cyclin-dependent kinase 2 is required for zebrafish embryogenesis', Molecular Biology of the Cell, vol. 23, no. 1, pp. 59-70. https://doi.org/10.1091/mbc.E11-06-0487
Chua, Jaime ; Loughlin, Elizabeth A. ; Gaur, Naseem A. ; SenBanerjee, Sucharita ; Jacob, Vinitha ; Monson, Christopher ; Kent, Brandon ; Oranu, Amanke ; Ding, Yuanying ; Ukomadu, Chinweike ; Sadler Edepli, Kirsten. / UHRF1 phosphorylation by cyclin A2/cyclin-dependent kinase 2 is required for zebrafish embryogenesis. In: Molecular Biology of the Cell. 2012 ; Vol. 23, No. 1. pp. 59-70.
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