NSD1; cell cycle; cytoskeleton; isoforms; neoplastic pathways; Histone-Lysine N-Methyltransferase; Protein Isoforms; NSD1 protein, human; Humans; Cell Division/genetics; Cell Line; Alternative Splicing; Stress Fibers/metabolism; Fibroblasts/metabolism; Actin Cytoskeleton/metabolism; Actin Cytoskeleton/genetics; Histone-Lysine N-Methyltransferase/genetics; Histone-Lysine N-Methyltransferase/metabolism; Protein Isoforms/genetics; Protein Isoforms/metabolism; Genetics; Genetics (clinical)
Abstract :
[en] Germline variants in the NSD1 gene are responsible for Sotos syndrome, while somatic variants promote neoplastic cell transformation. Our previous studies revealed three alternative RNA isoforms of NSD1 present in fibroblast cell lines (FBs): the canonical full transcript and 2 alternative transcripts, termed AT2 (NSD1 Δ5Δ7) and AT3 (NSD1 Δ19-23 at the 5' end). The precise molecular pathways affected by each specific isoform of NSD1 are uncharacterized to date. To elucidate the role of these isoforms, their expression was suppressed by siRNA knockdown in FBs and protein expression and transcriptome data was explored. We demonstrate that one gene target of NSD1 isoform AT2 is ARP3 actin-related protein 3 homolog B (ACTR3B). We show that loss of both canonical NSD1 and AT2 isoforms impaired the ability of fibroblasts to regulate the actin cytoskeleton, and we observed that this caused selective loss of stress fibers. Our findings provide novel insights into NSD1 function by distinguishing isoform function and demonstrating an essential role of NSD1 in regulating the actin cytoskeleton and stress fiber formation in fibroblasts.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Conteduca, Giuseppina ; Biotherapy Unit, IRCCS San Martino, 16132 Genoa, Italy
Baldo, Chiara ; Laboratory of Human Genetics, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy
Arado, Alessia; Laboratory of Human Genetics, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy
Testa, Barbara; Laboratory of Human Genetics, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy
Wagner, Ryan T ; Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 55905, USA
Robertson, Keith D ; Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, Rochester, MN 55905, USA
Dequiedt, Franck ; Université de Liège - ULiège > Département des sciences de la vie > Génétique et biologie moléculaires animales
Fitzsimmons, Lane; Renaissance School of Medicine, Stony Brook University, Stony Brook, NY 11794, USA
Malacarne, Michela; Laboratory of Human Genetics, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy
Filaci, Gilberto ; Biotherapy Unit, IRCCS San Martino, 16132 Genoa, Italy ; Department of Internal Medicine (DIMI), University of Genoa, 16132 Genoa, Italy
Coviello, Domenico A ; Laboratory of Human Genetics, IRCCS Istituto Giannina Gaslini, 16147 Genoa, Italy
Language :
English
Title :
Impact of NSD1 Alternative Transcripts in Actin Filament Formation and Cellular Division Pathways in Fibroblasts.
Publication date :
24 August 2024
Journal title :
Genes
ISSN :
2073-4425
Publisher :
Multidisciplinary Digital Publishing Institute (MDPI), Switzerland
ASSI Gulliver Fondazione di Sardegna Italian Ministry of Health
Funding text :
This research was funded by the AssiGulliver Italian Association; Fondazione Sardegna; Banca d’Italia, Italian Ministry of Health, Ricerca Corrente 2023 RRC-2023-23683432 entitled Allestimento di un modello cellulare in vitro per analizzare la componente molecolare responsabile della Sindrome di Sotos in previsione dell’individuazione di terapie innovative, 5M-2020-23682542, 5xmille grant from Italian Ministry of Health.
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