Neurofibromin 1 is a miRNA target in neurons.
Mutations of the neurofibromin 1 gene cause neurofibromatosis type 1, a disease in which learning and behavioral abnormalities are common. The disease is completely penetrant but shows variable phenotypic expression in patients. The repertoire of regulatory interactions utilized by neurons to control neurofibromin 1 expression is poorly understood. Here, we examined the contribution of microRNAs into this regulatory network. Using reporter assays, we provided evidence that miR-128 and to a lesser extent miR-137 and miR-103 reduced neurofibromin 1 reporter levels through specific binding to Nf1 3'-UTR. Mutations in all three predicted binding sites eliminated the reporter response. MiR-128 and miR-137, unlike miR-103 that showed a more ubiquitous expression, were predominantly expressed in brain with a distribution that resembled neurofibromin 1 expression in different tissues as well as during the course of neuronal development. In the nervous system, all three microRNAs showed highest expression in neurons and least in Schwann cells and astrocytes. Overexpression of miR-128 alone or with miR-103 and miR-137 significantly reduced endogenous neurofibromin 1 protein levels, while antisense inhibition of these microRNAs enhanced translation of endogenous neurofibromin 1 and reporter in primary cultures of hippocampal neurons. These findings revealed a significant additional mechanism by which neurofibromin 1 is regulated in neurons and implicated new candidates for the treatment of multifarious neurofibromatosis type 1 cognitive symptoms.
Molecular and Cellular Neurobiology Lab, Center of Neurosciences and Social Psychiatry, Biomedical Research Foundation, Academy of Athens, Athens, Greece.
Maria
M
Paschou
Epaminondas
E
Doxakis
D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D001253:Astrocytes / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D002478:Cells, Cultured; D006801:Humans; D007150:Immunohistochemistry; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D035683:MicroRNAs / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D025542:Neurofibromin 1 / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D012342:RNA, Small Nuclear / Q000378:metabolism
D020413:3' Untranslated Regions; C555364:MIRN103 microRNA, mouse; C551799:MIRN137 microRNA, mouse; D035683:MicroRNAs; C550324:Mirn128 microRNA, mouse; D025542:Neurofibromin 1; D012342:RNA, Small Nuclear; C405486:U6 small nuclear RNA