Kaufman oculocerebrofacial syndrome (KOS) is a severe autosomal recessive disorder characterized by intellectual disability, developmental delays, microcephaly, and characteristic dysmorphisms. Biallelic mutations of UBE3B, encoding for a ubiquitin ligase E3B are causative for KOS. In this report, we characterize neuronal functions of its murine ortholog Ube3b and show that Ube3b regulates dendritic branching in a cell-autonomous manner. Moreover, Ube3b knockout (KO) neurons exhibit increased density and aberrant morphology of dendritic spines, altered synaptic physiology, and changes in hippocampal circuit activity. Dorsal forebrain-specific Ube3b KO animals show impaired spatial learning, altered social interactions, and repetitive behaviors. We further demonstrate that Ube3b ubiquitinates the catalytic γ-subunit of calcineurin, Ppp3cc, the overexpression of which phenocopies Ube3b loss with regard to dendritic spine density. This work provides insights into the molecular pathologies underlying intellectual disability-like phenotypes in a genetically engineered mouse model.
This is a preview of subscription content, access via your institution
Subscribe to Journal
Get full journal access for 1 year
only $9.92 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Tønnesen J, Nägerl UV. Dendritic spines as tunable regulators of synaptic signals. Front Psychiatry. 2016;7:101.
Ambrozkiewicz MC, Kawabe H. HECT-type E3 ubiquitin ligases in nerve cell development and synapse physiology. FEBS Lett. 2015;589:1635–43.
Kawabe H, Brose N. The role of ubiquitylation in nerve cell development. Nat Rev Neurosci. 2011;12:251–68.
Hershko A, Heller H, Elias S, Ciechanover A. Components of ubiquitin-protein ligase system. Resolution, affinity purification, and role in protein breakdown. J Biol Chem. 1983;258:8206–14.
Kaiser SE, Riley BE, Shaler TA, Trevino RS, Becker CH, Schulman H, et al. Protein standard absolute quantification (PSAQ) method for the measurement of cellular ubiquitin pools. Nat Methods. 2011;8:691–6.
Hicke L, Riezman H. Ubiquitination of a yeast plasma membrane receptor signals its ligand-stimulated endocytosis. Cell. 1996;84:277–87.
Rotin D, Kumar S. Physiological functions of the HECT family of ubiquitin ligases. Nat Rev Mol Cell Biol. 2009;10:398–409.
Huibregtse JM, Scheffner M, Beaudenon S, Howley PM. A family of proteins structurally and functionally related to the E6-AP ubiquitin-protein ligase. Proc Natl Acad Sci USA. 1995;92:2563–7.
Ambrozkiewicz MC, Schwark M, Kishimoto-Suga M, Borisova E, Hori K, Salazar-Lázaro A, et al. Polarity acquisition in cortical neurons is driven by synergistic action of Sox9-regulated Wwp1 and Wwp2 E3 ubiquitin ligases and intronic miR-140. Neuron. 2018;100:1097–1115.e15.
Hsia H-E, Kumar R, Luca R, Takeda M, Courchet J, Nakashima J, et al. Ubiquitin E3 ligase Nedd4-1 acts as a downstream target of PI3K/PTEN-mTORC1 signaling to promote neurite growth. Proc Natl Acad Sci USA. 2014;111:13205–10.
Kawabe H, Neeb A, Dimova K, Young SM, Takeda M, Katsurabayashi S, et al. Regulation of Rap2A by the ubiquitin ligase Nedd4-1 controls neurite development. Neuron. 2010;65:358–72.
Basel-Vanagaite L, Dallapiccola B, Ramirez-Solis R, Segref A, Thiele H, Edwards A, et al. Deficiency for the ubiquitin ligase UBE3B in a blepharophimosis-ptosis-intellectual-disability syndrome. Am J Hum Genet. 2012;91:998–1010.
Basel-Vanagaite L, Yilmaz R, Tang S, Reuter MS, Rahner N, Grange DK, et al. Expanding the clinical and mutational spectrum of Kaufman oculocerebrofacial syndrome with biallelic UBE3B mutations. Hum Genet. 2014;133:939–49.
Flex E, Ciolfi A, Caputo V, Fodale V, Leoni C, Melis D, et al. Loss of function of the E3 ubiquitin-protein ligase UBE3B causes Kaufman oculocerebrofacial syndrome. J Med Genet. 2013;50:493–9.
Pedurupillay CRJ, Barøy T, Holmgren A, Blomhoff A, Vigeland MD, Sheng Y, et al. Kaufman oculocerebrofacial syndrome in sisters with novel compound heterozygous mutation in UBE3B. Am J Med Genet A. 2015;167A:657–63.
Chahrour MH, Yu TW, Lim ET, Ataman B, Coulter ME, Hill RS, et al. Whole-exome sequencing and homozygosity analysis implicate depolarization-regulated neuronal genes in autism. PLoS Genet. 2012;8:e1002635.
Svilar D, Dyavaiah M, Brown AR, Tang J, Li J, McDonald PR, et al. Alkylation sensitivity screens reveal a conserved cross-species functionome. Mol Cancer Res MCR. 2012;10:1580–96.
Braganza A, Li J, Zeng X, Yates NA, Dey NB, Andrews J, et al. UBE3B is a calmodulin-regulated, mitochondrion-associated E3 Ubiquitin Ligase. J Biol Chem. 2017;292:2470–84.
Cheon S, Kaur K, Nijem N, Tuncay IO, Kumar P, Dean M, et al. The ubiquitin ligase UBE3B, disrupted in intellectual disability and absent speech, regulates metabolic pathways by targeting BCKDK. Proc Natl Acad Sci. 2019;116:3662–7. https://doi.org/10.1073/pnas.1818751116.
Gorski JA, Talley T, Qiu M, Puelles L, Rubenstein JLR, Jones KR. Cortical excitatory neurons and glia, but not GABAergic neurons, are produced in the Emx1-expressing lineage. J Neurosci J Soc Neurosci. 2002;22:6309–14.
Lakso M, Pichel JG, Gorman JR, Sauer B, Okamoto Y, Lee E, et al. Efficient in vivo manipulation of mouse genomic sequences at the zygote stage. Proc Natl Acad Sci USA. 1996;93:5860–5.
Thomas KR, Capecchi MR. Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell. 1987;51:503–12.
Koch S, Mueller S, Foddis M, Bienert T, von Elverfeldt D, Knab F, et al. Atlas registration for edema-corrected MRI lesion volume in mouse stroke models. J Cereb Blood Flow Metab. 2019;39:313–23.
Zaqout S, Kaindl AM. Golgi–Cox staining step by step. Front Neuroanat. 2016;10:38.
Dere E, Dahm L, Lu D, Hammerschmidt K, Ju A, Tantra M, et al. Heterozygous ambra1 deficiency in mice: a genetic trait with autism-like behavior restricted to the female gender. Front Behav Neurosci. 2014;8:181.
Netrakanti PR, Cooper BH, Dere E, Poggi G, Winkler D, Brose N, et al. Fast cerebellar reflex circuitry requires synaptic vesicle priming by munc13-3. Cerebellum Lond Engl. 2015;14:264–83.
Telley L, Agirman G, Prados J, Amberg N, Fièvre S, Oberst P, et al. Temporal patterning of apical progenitors and their daughter neurons in the developing neocortex. Science. 2019;364. pii: eaav2522. https://doi.org/10.1126/science.aav2522.
Song JY, Ichtchenko K, Südhof TC, Brose N. Neuroligin 1 is a postsynaptic cell-adhesion molecule of excitatory synapses. Proc Natl Acad Sci USA. 1999;96:1100–5.
Coley AA, Gao W-J. PSD95: A synaptic protein implicated in schizophrenia or autism? Prog Neuropsychopharmacol Biol Psychiatry. 2018;82:187–94.
Nakanishi M, Nomura J, Ji X, Tamada K, Arai T, Takahashi E, et al. Functional significance of rare neuroligin 1 variants found in autism. PLoS Genet. 2017;13:e1006940.
Wang Q, Moore MJ, Adelmant G, Marto JA, Silver PA. PQBP1, a factor linked to intellectual disability, affects alternative splicing associated with neurite outgrowth. Genes Dev. 2013;27:615–26.
Guo H, Hong S, Jin XL, Chen RS, Avasthi PP, Tu YT, et al. Specificity and efficiency of Cre-mediated recombination in Emx1-Cre knock-in mice. Biochem Biophys Res Commun. 2000;273:661–5.
Koleske AJ. Molecular mechanisms of dendrite stability. Nat Rev Neurosci. 2013;14:536–50.
Rosário M, Schuster S, Jüttner R, Parthasarathy S, Tarabykin V, Birchmeier W. Neocortical dendritic complexity is controlled during development by NOMA-GAP-dependent inhibition of Cdc42 and activation of cofilin. Genes Dev. 2012;26:1743–57.
Dindot SV, Antalffy BA, Bhattacharjee MB, Beaudet AL. The Angelman syndrome ubiquitin ligase localizes to the synapse and nucleus, and maternal deficiency results in abnormal dendritic spine morphology. Hum Mol Genet. 2008;17:111–8.
Saito T. In vivo electroporation in the embryonic mouse central nervous system. Nat Protoc. 2006;1:1552–8.
Bekkers JM, Stevens CF. Excitatory and inhibitory autaptic currents in isolated hippocampal neurons maintained in cell culture. Proc Natl Acad Sci USA. 1991;88:7834–8.
Ripamonti S, Ambrozkiewicz MC, Guzzi F, Gravati M, Biella G, Bormuth I, et al. Transient oxytocin signaling primes the development and function of excitatory hippocampal neurons. ELife. 2017;6. pii: e22466. https://doi.org/10.7554/eLife.22466.
Chau V, Tobias JW, Bachmair A, Marriott D, Ecker DJ, Gonda DK, et al. A multiubiquitin chain is confined to specific lysine in a targeted short-lived protein. Science. 1989;243:1576–83.
Duncan LM, Piper S, Dodd RB, Saville MK, Sanderson CM, Luzio JP, et al. Lysine-63-linked ubiquitination is required for endolysosomal degradation of class I molecules. EMBO J. 2006;25:1635–45.
Grice GL, Nathan JA. The recognition of ubiquitinated proteins by the proteasome. Cell Mol Life Sci CMLS. 2016;73:3497–506.
Gong TW, Hegeman AD, Shin JJ, Adler HJ, Raphael Y, Lomax MI. Identification of genes expressed after noise exposure in the chick basilar papilla. Hear Res. 1996;96:20–32.
Mazumder MK, Paul R, Bhattacharya P, Borah A. Neurological sequel of chronic kidney disease: from diminished Acetylcholinesterase activity to mitochondrial dysfunctions, oxidative stress and inflammation in mice brain. Sci Rep. 2019;9:3097.
Vanholder R, De Smet R, Glorieux G, Argilés A, Baurmeister U, Brunet P, et al. Review on uremic toxins: classification, concentration, and interindividual variability. Kidney Int. 2003;63:1934–43.
Ramakers GJ. Rho proteins and the cellular mechanisms of mental retardation. Am J Med Genet. 2000;94:367–71.
Dierssen M, Ramakers GJA. Dendritic pathology in mental retardation: from molecular genetics to neurobiology. Genes Brain Behav. 2006;5:48–60.
Engert F, Bonhoeffer T. Dendritic spine changes associated with hippocampal long-term synaptic plasticity. Nature 1999;399:66–70.
Noguchi J, Matsuzaki M, Ellis-Davies GCR, Kasai H. Spine-neck geometry determines NMDA receptor-dependent Ca2+ signaling in dendrites. Neuron 2005;46:609–22.
Verma V, Paul A, Amrapali Vishwanath A, Vaidya B, Clement JP. Understanding intellectual disability and autism spectrum disorders from common mouse models: synapses to behaviour. Open Biol. 2019;9:180265.
Comery TA, Harris JB, Willems PJ, Oostra BA, Irwin SA, Weiler IJ, et al. Abnormal dendritic spines in fragile X knockout mice: maturation and pruning deficits. Proc Natl Acad Sci. 1997;94:5401–4.
Csicsvari J, Jamieson B, Wise KD, Buzsáki G. Mechanisms of gamma oscillations in the hippocampus of the behaving rat. Neuron. 2003;37:311–22.
Hitti FL, Siegelbaum SA. The hippocampal CA2 region is essential for social memory. Nature. 2014;508:88–92.
Silverman JL, Yang M, Lord C, Crawley JN. Behavioural phenotyping assays for mouse models of autism. Nat Rev Neurosci. 2010;11:490–502.
Yilmaz R, Szakszon K, Altmann A, Altunoglu U, Senturk L, McGuire M, et al. Kaufman oculocerebrofacial syndrome: novel UBE3B mutations and clinical features in four unrelated patients. Am J Med Genet A. 2018;176:187–93.
American Psychiatric Association. diagnostic and statistical manual of mental disorders. 5th ed. American Psychiatric Association; 2013. https://www.psychiatry.org/psychiatrists/practice/dsm.
Sutton LP, Orlandi C, Song C, Oh WC, Muntean BS, Xie K, et al. Orphan receptor GPR158 controls stress-induced depression. ELife. 2018;7. pii: e33273. https://doi.org/10.7554/eLife.33273.
van Diepen MT, Parsons M, Downes CP, Leslie NR, Hindges R, Eickholt BJ. MyosinV controls PTEN function and neuronal cell size. Nat Cell Biol. 2009;11:1191–6.
Konietzny A, González-Gallego J, Bär J, Perez-Alvarez A, Drakew A, Demmers JAA, et al. Myosin V regulates synaptopodin clustering and localization in the dendrites of hippocampal neurons. J Cell Sci. 2019;132. pii: jcs230177. https://doi.org/10.1242/jcs.230177.
Belvindrah R, Natarajan K, Shabajee P, Bruel-Jungerman E, Bernard J, Goutierre M, et al. Mutation of the α-tubulin Tuba1a leads to straighter microtubules and perturbs neuronal migration. J Cell Biol. 2017;216:2443–61.
Chandran V, Coppola G, Nawabi H, Omura T, Versano R, Huebner EA, et al. A systems-level analysis of the peripheral nerve intrinsic axonal growth program. Neuron. 2016;89:956–70.
Li M, Wen H, Yan Z, Ding T, Long L, Qin H, et al. Temporal-spatial expression of ENOLASE after acute spinal cord injury in adult rats. Neurosci Res. 2014;79:76–82.
Lin L, Sun W, Kung F, Dell’Acqua ML, Hoffman DA. AKAP79/150 impacts intrinsic excitability of hippocampal neurons through phospho-regulation of A-type K+ channel trafficking. J Neurosci J Soc Neurosci. 2011;31:1323–32.
Alagarsamy S, Saugstad J, Warren L, Mansuy IM, Gereau RW, Conn PJ. NMDA-induced potentiation of mGluR5 is mediated by activation of protein phosphatase 2B/calcineurin. Neuropharmacology. 2005;49:135–45.
Horiuchi Y, Ishiguro H, Koga M, Inada T, Iwata N, Ozaki N, et al. Support for association of the PPP3CC gene with schizophrenia. Mol Psychiatry. 2007;12:891–3.
Miyakawa T, Leiter LM, Gerber DJ, Gainetdinov RR, Sotnikova TD, Zeng H, et al. Conditional calcineurin knockout mice exhibit multiple abnormal behaviors related to schizophrenia. Proc Natl Acad Sci. 2003;100:8987–92.
This work was supported by the German Research Foundation (SPP1365/KA3423/1-1 and KA3423/3-1, HK; DFG TA 303/4-2, VT), and the Russian Scientific Foundation (19-14-00345, VT), JSPS KAKENHI Grant Numbers 15K21769 (HK), The Mother and Child Health Foundation (HK), the Uehara Memorial Foundation (HK), and the Fritz Thyssen Foundation (HK). Funding to SM and PBS was provided by the German Federal Ministry of Education and Research (BMBF, Center for Stroke Research Berlin 01EO1301), the BMBF under the ERA-NET NEURON scheme (01EW1811), and the German Research Foundation (DFG, Project 428869206 and EXC NeuroCure). We would like to acknowledge Nils Brose for his help with project administration and insightful remarks regarding the conceptualization. We thank Fritz Benseler, Klaus-Peter Hellman, Bernd Hesse-Niessen, Ivonne Thanhäuser, Dayana Warnecke, Christiane Harenberg, Maik Schlieper, Dörte Hesse, Stefan Paul Koch, Jutta Schüler, Olaf Ninnemann, Claudia Pallasch, Rike Dannenberg, and Denis Lajkó and animal facilities of Max Planck Institute of Experimental Medicine and of Charité University Hospital. We acknowledge Alex Knebel, Thimo Kurz, Carlos Henrique Vieira e Vieira, Matthias Selbach, Hannelore Ehrenreich, Ahmed Mansouri, and Judith Stegmüller for their feedback.
Conflict of interest
The authors declare that they have no conflict of interest.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Ambrozkiewicz, M.C., Borisova, E., Schwark, M. et al. The murine ortholog of Kaufman oculocerebrofacial syndrome protein Ube3b regulates synapse number by ubiquitinating Ppp3cc. Mol Psychiatry 26, 1980–1995 (2021). https://doi.org/10.1038/s41380-020-0714-8