title: Ubiquitin C-terminal hydrolase L1 interacts with choline transporter in cholinergic cells.
abstract: Ubiquitin carboxyl-terminal hydrolase L1 (UCHL1) is a deubiquitinating enzyme, which is highly expressed in neuronal cells. Previous studies have indicated that UCHL1 is involved in cognitive function, neurodegenerative diseases, and neuromuscular junction development. Acetylcholine (Ach) is a critical neurotransmitter in these functions. Yet, the effect of UCHL1 on the cholinergic system has not been reported. In this study, using a cholinergic neuronal cell line, SN56, as an invitro model, we detected the physical interaction of UCHL1 and high affinity choline transporter (CHT), which is a key protein regulating Ach re-synthesis. Reduction of UCHL1 by siRNA gene knockdown significantly increased poly-ubiquitinated CHT and decreased native CHT protein level, but did not affect CHT mRNA expression. Biotinylation assay showed that UCHL1 is localized only in the cytosol of the cells and that the gene knockdown of UCHL1 significantly reduced cytosolic CHT but had no significant effect on membrane CHT level. These data provide novel and potentially important evidence that UCHL1 may play a role in the regulation of cholinergic function by affecting CHT ubiquitination and degradation.
authors: Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South Dakota, Vermillion, SD, 57069, USA.
authors: Sigurd
authors: S
authors: Hartnett
authors: Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South Dakota, Vermillion, SD, 57069, USA.
authors: Fan
authors: F
authors: Zhang
authors: Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South Dakota, Vermillion, SD, 57069, USA.
authors: Allison
authors: A
authors: Abitz
authors: Division of Basic Biomedical Sciences, Sanford School of Medicine, The University of South Dakota, Vermillion, SD, 57069, USA. Electronic address: yli01@usd.edu.
authors: Yifan
authors: Y
authors: Li
mesh_terms: D000818:Animals; D002460:Cell Line; D059329:Cholinergic Neurons / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000378:metabolism*; D051379:Mice; D043222:Ubiquitin Thiolesterase / Q000378:metabolism*
chemical_list: D026901:Membrane Transport Proteins; C065637:choline transporter; D043222:Ubiquitin Thiolesterase
keywords: Acetylcholine; CHT; SN56 cells; UCHL1; Ubiquitination