| Annotated protein: | Presenilin-1 (PS-1) (EC 3.4.23.-) (Protein S182) [Cleaved into: Presenilin-1 NTF subunit; Presenilin-1 CTF subunit; Presenilin-1 CTF12 (PS1-CTF12)]. Gene symbol: PSEN1. Taxonomy: Mus musculus (Mouse). Uniprot ID: P49769 |
| antibody wiki: | |
| SynGO gene info: | SynGO data @ PSEN1 |
| Ontology domain: | Biological Process |
| SynGO term: | regulation of postsynapse organization (GO:0099175) |
| Synapse type(s): | hippocampus, glutamatergic |
| Annotated paper: | Suzuki K, et al. "Activity-dependent proteolytic cleavage of neuroligin-1" Neuron. 2012 Oct 18;76(2):410-22 PMID:23083742 |
| Figure(s): | Fig. 2, 3, 5, 8 |
| Annotation description: | 20/3/2018 Pim Note: the annotation for Presenilin-1 and Presenilin-2 is correlational. 1] Presenilin-1/2 are shown to cleave NGL1 in non-neuronal cells, and 2] the truncated NGL1 version is shown to lack synaptogenic activity. I decided that this information together with the data on ADAM10, which is a known 'shedase' for gamma-secretase, is still convincing enough to ward an annotation. Background: This paper shows that NLG1 is proteolytically cleaved in a two step process by the ADAM10 and gamma-secretase protease in sequence. Cleavage by ADAM10 ratains the synaptogenic activity of NLG1, but is needed for gamma-sectretase to complete the job and remove NLG1 activity. Fig.2: "Proteolytic Processing of NLG1 in Presenilin or ADAM Knockout Cells" - "Next we analyzed the metabolism of NLGs in mouse embryonic fibroblasts from Psen1-/-/Psen2-/- double knockout mice (DKO cells), which completely lacks the γ-secretase activity (Herreman et al., 2000). Accumulation of NLG-CTFs was observed upon the overexpression of hemagglutinin (HA)-tagged NLGs in DKO cells (Figure 2A). However, the levels of the accumulated NLG-CTFs were significantly reduced by the coexpression of human PS1, indicating that γ-secretase activity is responsible for the processing of NLG-CTFs" - "To test whether ADAM10 is involved in the processing of NLGs, we overexpressed HA-tagged NLG1 or NLG2 in murine embryonic fibroblasts (MEFs) obtained from ADAM10 knockout (Adam10-/-) or heterozygous (Adam10+/-) mice (Figure 2B) (Hartmann et al., 2002). In Adam10-/- MEF, the generation of sNLG1 was significantly reduced. In contrast, no change in NLG1 processing was observed in MEFs obtained from knockout mice of other ADAMs (i.e., Adam8-/-, Adam17-/-, Adam19-/-, Adam9-/-;Adam12-/-;Adam15-/- [TKO]) " Fig.3: "Pharmacological and Genetic Analyses of NLG1 Shedding in Neurons - "To further examine the role of ADAM10 in the processing of endogenous NLG1, we treated rat primary neurons obtained from E18 pups with INCB3619, a known ADAM10/17 inhibitor (Witters et al., 2008). INCB3619 abolished the secretion of sNLG1 in a similar manner to that of sAPPα, the latter being generated by ADAM10 (IC50: 1.6 μM) " - "Moreover, overexpression of ADAM10 in mouse primary neurons increased the production of sNLG1 (Figure S2B). Lastly, to demonstrate the physiological significance of ADAM10 in NLG1 processing in vivo, we incubated the brain microsome from postnatal day (P) 18 neuron-specific conditional ADAM10 knockout mice (Adam10flox/flox; CamKII-Cre) (Figure 3I). Notably, the levels of NLG1-FL were increased in the microsome fractions from brains of Adam10 conditional knockout mice, whereas sNLG1 production was significantly decreased (Figures 3J and 3K). Taken together, we concluded that the major physiological sheddase of NLG1 in brain is ADAM10." Fig.5: "Activity-Dependent Shedding of NLG1 in Primary Neurons" - "To investigate the effect of synaptic activity on NLG1 processing, we treated rat primary neuronal culture at day in vitro (DIV) 11 with a set of compounds. Fifteen minute treatments with glutamate or NMDA significantly increased the sNLG1 level in the conditioned media, which was abolished by addition of NMDA receptor antagonists (i.e., D-AP5 and MK-801)" Fig.8: "Effect of Truncated NLG1 on Spine Formation" - "To analyze the functional impact of NLG1 processing on its spinogenic activity, we overexpressed NLG1 and its derivatives in dentate granule cells of the organotypic hippocampal slice culture obtained from P6 rat, in which local-circuit synaptic interactions are preserved. Overexpression of NLG1-FL significantly increased the spine density at the apical dendrites of granule cells. " - "Unexpectedly, NLG1-ΔE increased the spine density at a similar level to NLG1-FL, suggesting that the NLG1-CTF lacking the ectodomain retains the spinogenic effect. However, NLG1-ICD failed to increase the spine density. Thus, the function of membrane-tethered form of NLG1-ICD (aka, NLG1-ΔE or NLG1-CTF) was abolished by liberation from the membrane by the γ-secretase cleavage and subsequent degradation. Finally, to directly test whether NLG1 shedding modulates the spinogenic function, we analyzed the dendritic spines of transfected rat hippocampal primary neurons obtained from E18 pups (Figure 8C). We transfected wild-type or PKQQ/AAAA mutant NLG1 together with green fluorescent protein (GFP) into primary neurons at DIV6 and fixed them at DIV20. The numbers of spines in neurons expressing wild-type NLG1 showed an increased trend compared to those in mock-transfected neurons, but not with a statistical significance. However, the spine density was significantly increased in neurons transfected with the mutant NLG1 (Figure 8D), suggesting that cleavage-deficient mutation enhanced the NLG1 function in primary neurons." |
| Evidence tracking, Biological System: | Intact tissue Cultured neurons Non-neuronal tissue |
| Evidence tracking, Protein Targeting: | Genetic transformation (eg; knockout, knockin, mutations) |
| Evidence tracking, Experiment Assay: | Confocal Western blot |
| Annotator(s): | Pim van Nierop (ORCID:0000-0003-0593-3443) Guus Smit (ORCID:0000-0002-2286-1587) Matthijs Verhage (ORCID:0000-0002-2514-0216) |
| Lab: | Department of Functional Genomics, Department of Molecular and Cellular Neurobiology, Center for Neurogenomics and Cognitive Research, Vrije Universiteit Amsterdam, 1081 HV Amsterdam, The Netherlands |
| SynGO annotation ID: | 2971 |
| Dataset release (version): | 20231201 |
| View annotation as GO-CAM model: |  |