title: UHRF1 phosphorylation by cyclin A2/cyclin-dependent kinase 2 is required for zebrafish embryogenesis.
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.
authors.affiliation: Division of Pediatric Hepatology, Department of Pediatrics, Mount Sinai School of Medicine, New York, NY 10029, USA.
authors.forename: Jaime
authors.initials: J
authors.lastname: Chu
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Loughlin
authors.forename: Naseem A
authors.initials: NA
authors.lastname: Gaur
authors.forename: Sucharita
authors.initials: S
authors.lastname: SenBanerjee
authors.forename: Vinitha
authors.initials: V
authors.lastname: Jacob
authors.forename: Christopher
authors.initials: C
authors.lastname: Monson
authors.forename: Brandon
authors.initials: B
authors.lastname: Kent
authors.forename: Amanke
authors.initials: A
authors.lastname: Oranu
authors.forename: Yuanying
authors.initials: Y
authors.lastname: Ding
authors.forename: Chinweike
authors.initials: C
authors.lastname: Ukomadu
authors.forename: Kirsten C
authors.initials: KC
authors.lastname: Sadler
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D016384:Consensus Sequence; D056751:Cyclin A2 / Q000235:genetics / Q000378:metabolism*; D051357:Cyclin-Dependent Kinase 2 / Q000378:metabolism*; D004625:Embryo, Nonmammalian / Q000033:anatomy & histology; D047108:Embryonic Development; D005775:Gastrula / Q000196:embryology; D018507:Gene Expression Regulation, Developmental; D055785:Gene Knockdown Techniques; D008969:Molecular Sequence Data; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D015534:Trans-Activators / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015027:Zebrafish / Q000196:embryology* / Q000235:genetics; D029961:Zebrafish Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D056751:Cyclin A2; D015534:Trans-Activators; C517793:Uhrf1 protein, zebrafish; D029961:Zebrafish Proteins; C503723:ccna2 protein, zebrafish; D051357:Cyclin-Dependent Kinase 2
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