The Networks of Genes Encoding Palmitoylated Proteins in Axonal and Synaptic Compartments Are Affected in PPT1 Overexpressing Neuronal-Like Cells
| Autor: | Francesca Griggio, Marzia Bianchi, Rosalba Carrozzo, Salvatore Benfatto, Massimo Delledonne, Alessandro Simonati, Filippo M. Santorelli, Arvydas Dapkunas, Stefano Doccini, Maciej Lalowski, Francesco Pezzini |
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| Přispěvatelé: | Medicum, Department of Biochemistry and Developmental Biology, University of Helsinki |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
congenital hereditary and neonatal diseases and abnormalities CLN1 disease PELIZAEUS-MERZBACHER-DISEASE NEFM GAP-43 Biology 3124 Neurology and psychiatry lcsh:RC321-571 Transcriptome 03 medical and health sciences Cellular and Molecular Neuroscience PPT1 0302 clinical medicine Palmitoylation THIOESTERASE 1 dysregulated genes HUMAN NEUROBLASTOMA-CELLS palmitoylation Gap-43 protein CEROID-LIPOFUSCINOSIS GROWTH-ASSOCIATED PROTEIN CLN1 GENE lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry Molecular Biology Original Research INTERMEDIATE-FILAMENTS 3112 Neurosciences Wild type SNAP25 INFANTILE TYPE Transfection Cell biology MICE Differentiated neuroblastoma cells Dysregulated genes Neuritogenesis RNA-seq 030104 developmental biology differentiated neuroblastoma cells neuritogenesis biology.protein 3111 Biomedicine 030217 neurology & neurosurgery Neuroscience |
| Zdroj: | Frontiers in Molecular Neuroscience, Vol 10 (2017) Frontiers in Molecular Neuroscience |
| ISSN: | 1662-5099 |
| DOI: | 10.3389/fnmol.2017.00266 |
| Popis: | CLN1 disease (OMIM # 256730) is an early childhood ceroid-lipofuscinosis associated with mutated CLN1, whose product Palmitoyl-Protein Thioesterase 1 (PPT1) is a lysosomal enzyme involved in the removal of palmitate residues from S-acylated proteins. In neurons, PPT1 expression is also linked to synaptic compartments. The aim of this study was to unravel molecular signatures connected to CLN1. We utilized SH-SY5Y neuroblastoma cells overexpressing wild type CLN1 (SH-p. wtCLN1) and five selected CLN1 patients' mutations. The cellular distribution of wtPPT1 was consistent with regular processing of endogenous protein, partially detected inside Lysosomal Associated Membrane Protein 2 (LAMP2) positive vesicles, while the mutants displayed more diffuse cytoplasmic pattern. Transcriptomic profiling revealed 802 differentially expressed genes (DEGs) in SH-p. wtCLN1 (as compared to empty-vector transfected cells), whereas the number of DEGs detected in the two mutants (p. L222P and p. M57Nfs * 45) was significantly lower. Bioinformatic scrutiny linked DEGs with neurite formation and neuronal transmission. Specifically, neuritogenesis and proliferation of neuronal processes were predicted to be hampered in the wtCLN1 overexpressing cell line, and these findings were corroborated by morphological investigations. Palmitoylation survey identified 113 palmitoylated protein-encoding genes in SH-p. wtCLN1, including 25 ones simultaneously assigned to axonal growth and synaptic compartments. A remarkable decrease in the expression of palmitoylated proteins, functionally related to axonal elongation (GAP43, CRMP1 and NEFM) and of the synaptic marker SNAP25, specifically in SH-p. wtCLN1 cells was confirmed by immunoblotting. Subsequent, bioinformatic network survey of DEGs assigned to the synaptic annotations linked 81 DEGs, including 23 ones encoding for palmitoylated proteins. Results obtained in this experimental setting outlined two affected functional modules (connected to the axonal and synaptic compartments), which can be associated with an altered gene dosage of wtCLN1. Moreover, these modules were interrelated with the pathological effects associated with loss of PPT1 function, similarly as observed in the Ppt1 knockout mice and patients with CLN1 disease. |
| Databáze: | OpenAIRE |
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