The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and Glycosylation

Nobukuni, Mariko; Mochizuki, Hideki; Okada, Satoshi; Kameyama, Nobuhisa; Tanaka, Anna; Yamamoto, Hikaru; Amano, Taku; Seki, Takahiro; Sakai, Norio

doi:10.1254/jphs.09195fp

Cited by 21 publications

(39 citation statements)

References 50 publications

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“…A band with a molecular size of approximately 63 kDa (arrowhead) was identified as the form of SERT that is modified by high-mannose-type oligosaccharides, which is the immaturely glycosylated SERT that is retained in the ER. Our previous study also demonstrated that bands with molecular sizes greater than 80 kDa correspond to oligomers of immaturely glycosylated SERT (18,37). As shown in Fig.…”

Section: -Pba Increased Maturely Glycosylated Sert and Decreased Immsupporting

confidence: 56%

“…Previously, we examined whether proteasome inhibition affects the uptake activity and membrane trafficking of SERT (18). We found that proteasome inhibition caused SERT to be retained in the ER, which resulted in ER stress due to impaired membrane trafficking.…”

Section: Effects Of the Chemical Chaperone 4-phenylbutylate On The Fumentioning

confidence: 99%

“…The plasmid is designated here as pFLAG-SERT. A plasmid that can express a mutated form of SERT that lacks the C-terminus was made as described previously (18). This plasmid is designated here as pFLAG-SERTCT.…”

Section: Generation Of Plasmids and Adenoviral Vectorsmentioning

confidence: 99%

“…Immunocytochemistry was carried out for FLAG-SERT-expressing HEK293 cells as previously described (18,32). In brief, the cells were washed in 1 ml PBS and fixed with 4% paraformaldehyde and 0.2% picric acid in 0.1 M phosphate-buffered saline (PBS, pH 7.4) for 30 min at RT, washed twice with 1 ml PBS, and then permeabilized with 500 l PBS supplemented with 0.3% Triton X-100 and 5% normal goat serum (NGS) for 15 min.…”

Section: Immunocytochemistry and Analysis Of Plasma Membrane Expressimentioning

confidence: 99%

“…The C-terminus of SERT is known to play an important role in SERT membrane trafficking (17,18). We previously reported that SERTCT, which has a dramatically reduced uptake activity, was retained in ER and induced ER stress (18).…”

Section: -Pba Restored the Uptake Activity Of The C-terminaldeleted mentioning

confidence: 99%

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Effects of the Chemical Chaperone 4-Phenylbutylate on the Function of the Serotonin Transporter (SERT) Expressed in COS-7 Cells

Fujiwara¹,

Yamamoto²,

Miyagi³

et al. 2013

J Pharmacol Sci

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Abstract. The serotonin transporter (SERT) is involved in various psychiatric disorders, including depression and autism. Recently, chemical chaperones have been focused as potential therapeutic drugs that can improve endoplasmic reticulum (ER) stress-related pathology. In this study, we used SERT-transfected COS-7 cells to investigate whether 4-phenylbutylate (4-PBA), a chemical chaperone, affects the membrane trafficking and uptake activity of SERT. Treatment with 4-PBA for 24 h dose-dependently increased the uptake activity of SERT. In accordance with increased SERT activity, the expression of maturely glycosylated SERT was increased, while the expression of immaturely glycosylated SERT was decreased. This finding suggests that 4-PBA increased the functional SERT with mature glycosylation via accelerating its folding and trafficking. 4-PBA also increased the activity of the C-terminus-deleted mutant SERT (SERTCT), which was stacked in the ER, and decreased SERTCT-induced ER stress, further supporting the idea that 4-PBA acts as a chemical chaperone for SERT. Imaging studies showed that fluorescencelabeled SERT was gradually and significantly translocated to the plasma membrane by 4-PBA. These results suggest that 4-PBA and related drugs can potentially affect serotonergic neural transmission by functioning as chaperones, thereby providing a novel therapeutic approach for SERT-related diseases.[Supplementary materials: available only at http://dx

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Section: -Pba Increased Maturely Glycosylated Sert and Decreased Immsupporting

confidence: 56%

Section: Effects Of the Chemical Chaperone 4-phenylbutylate On The Fumentioning

confidence: 99%

Section: Generation Of Plasmids and Adenoviral Vectorsmentioning

confidence: 99%

Section: Immunocytochemistry and Analysis Of Plasma Membrane Expressimentioning

confidence: 99%

Section: -Pba Restored the Uptake Activity Of The C-terminaldeleted mentioning

confidence: 99%

See 3 more Smart Citations

Effects of the Chemical Chaperone 4-Phenylbutylate on the Function of the Serotonin Transporter (SERT) Expressed in COS-7 Cells

Fujiwara¹,

Yamamoto²,

Miyagi³

et al. 2013

J Pharmacol Sci

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RETRACTED: Placental serotonin systems in pregnancy metabolic complications associated with maternal obesity and gestational diabetes mellitus

Murthi

Vaillancourt

2020

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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Role of the E3 ubiquitin ligase HRD1 in the regulation of serotonin transporter function

Taguchi

Kaneko

Motoike

et al. 2021

Biochemical and Biophysical Research Communications

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To elucidate the regulation of serotonin transporter (SERT) function via its membrane trafficking, we investigated the involvement of the ubiquitin E3 ligase HRD1 (HMG-CoA reductase degradation protein), which participates in endoplasmic reticulum (ER)-associated degradation (ERAD), in the functional regulation of SERT. Cells transiently expressing wild-type SERT or a SERT C-terminal deletion mutant (SERTDCT), a SERT protein predicted to be misfolded, were used for experiments. Studies using HRD1overexpressing or HRD1-knockdown cells demonstrated that HRD1 is involved in SERT proteolysis. Overexpression of HRD1 promoted SERT ubiquitination, the effect of which was augmented by treatment with the proteasome inhibitor MG132. Immunoprecipitation studies revealed that HRD1 interacts with SERT in the presence of MG132. In addition, HRD1 was intracellularly colocalized with SERT, especially with aggregates of SERTDCT in the ER. HRD1 also affected SERT uptake activity in accordance with the expression levels of the SERT protein. These results suggest that HRD1 contributes to the membrane trafficking and functional regulation of SERT through its involvement in ERAD-mediated SERT degradation.

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The C-Terminal Region of Serotonin Transporter Is Important for Its Trafficking and Glycosylation

Cited by 21 publications

References 50 publications

Effects of the Chemical Chaperone 4-Phenylbutylate on the Function of the Serotonin Transporter (SERT) Expressed in COS-7 Cells

Effects of the Chemical Chaperone 4-Phenylbutylate on the Function of the Serotonin Transporter (SERT) Expressed in COS-7 Cells

RETRACTED: Placental serotonin systems in pregnancy metabolic complications associated with maternal obesity and gestational diabetes mellitus

Role of the E3 ubiquitin ligase HRD1 in the regulation of serotonin transporter function

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