Determination of topological structure of ARL6ip1 in cells: Identification of the essential binding region of ARL6ip1 for conophylline

Kuroda, Masahiro; Funasaki, Shintaro; Saitoh, Tohru; Sasazawa, Yukiko; Nishiyama, Shigeru; Umezawa, Kazuo; Simizu, Siro

doi:10.1016/j.febslet.2013.09.017

Cited by 15 publications

(9 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The topology of DPY19L3 was determined by redox‐sensitive luciferase assay using Gaussia luciferase (Gluc) . Gluc is a redox reporter that requires an oxidative environment to form disulfide bonds that are essential for its activity .…”

Section: Resultsmentioning

confidence: 99%

Topological analysis of DPY19L3, a human C‐mannosyltransferase

et al. 2018

Self Cite

View full text Add to dashboard Cite

C-mannosylation is a rare type of protein glycosylation, the functions and mechanisms of which remain unclear. Recently, we identified DPY19L3 as a C-mannosyltransferase of R-spondin1 in human cells. DPY19L3 is predicted to be a multipass transmembrane protein that localizes to the endoplasmic reticulum (ER); however, its structure is undetermined. In this study, we propose a topological structure of DPY19L3 by in silico analysis and experimental methods such as redox-sensitive luciferase assay and introduction of N-glycosylation sites, suggesting that DPY19L3 comprises 11 transmembrane regions and two re-entrant loops with the N- and C-terminal ends facing the cytoplasm and ER lumen, respectively. Furthermore, DPY19L3 has four predicted N-glycosylation sites, and we have demonstrated that DPY19L3 is N-glycosylated at Asn and Asn but not Asn and Asn , supporting our topological model. By mass spectrometry, we measured the C-mannosyltransferase activity of N-glycosylation-defective mutants of DPY19L3 and isoform2, a splice variant, which lacks the C-terminal luminal region of DPY19L3. Isoform2 does not possess C-mannosyltransferase activity, indicating the importance of the C-terminal region; however, N-glycosylations of DPY19L3 do not have any roles for its enzymatic activity. These novel findings on DPY19L3 provide important insights into the mechanism of C-mannosylation.

show abstract

Section: Resultsmentioning

confidence: 99%

Topological analysis of DPY19L3, a human C‐mannosyltransferase

et al. 2018

Self Cite

View full text Add to dashboard Cite

show abstract

“…2 A). As Arl6ip1 is an ER-integrated protein that associates with addicsin under normal conditions [5] , [17] , it is suitable for examining the change of intracellular addicsin localization upon PTZ exposure. Indeed, addicsin immunoreactivity (addicsin-IR) was shown to be predominantly co-localized with Arl6ip1 immunoreactivity (Arl6ip1-IR) in the ER under normal conditions ( Fig.…”

Section: Resultsmentioning

confidence: 99%

Pentylenetetrazol modulates redox system by inducing addicsin translocation from endoplasmic reticulum to plasma membrane in NG108-15 cells

Ikemoto

Murasawa

Wang

2017

Biochemistry and Biophysics Reports

View full text Add to dashboard Cite

Addicsin (Arl6ip5) is a multifunctional physiological and pathophysiological regulator that exerts its effects by readily forming homo- and hetero-complexes with various functional factors. In particular, addicsin acts as a negative modulator of neural glutamate transporter excitatory amino acid carrier 1 (EAAC1) and participates in the regulation of intracellular glutathione (GSH) content by negatively modulating EAAC1-mediated cysteine and glutamate uptake. Addicsin is considered to play a crucial role in the onset of neurodegenerative diseases including epilepsy. However, the molecular dynamics of addicsin remains largely unknown. Here, we report the dynamics of addicsin in NG108-15 cells upon exposure to pentylenetetrazol (PTZ), a representative epileptogenic agent acting on the gamma-Aminobutyric acid A (GABAA) receptor. Fluorescent immunostaining analysis demonstrated that addicsin drastically changed its localization from the endoplasmic reticulum (ER) to the plasma membrane within 1 h of PTZ exposure in a dose-dependent manner. Moreover, addicsin was co-localized with the plasma membrane markers EAAC1 and Na+/K+ ATPase alpha-3 upon PTZ stimulation. This translocation was significantly inhibited by a non-competitive GABAA receptor antagonist, picrotoxin, but not by a competitive GABAA receptor antagonist, bicuculline. Furthermore, lactate dehydrogenase (LDH) assay and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay showed that PTZ-induced addicsin translocation was accompanied by a decrease of radical-scavenging activity and an increase of cytotoxicity in a PTZ dose-dependent manner. These findings suggest that PTZ induces the translocation of addicsin from the ER to the plasma membrane and modulates the redox system by regulating EAAC1-mediated GSH synthesis, which leads to the activation of cell death signaling.

show abstract

“…On the other hand, ARL6IP1 and PSMC6 have been associated with insulin synthesis and pathology of diabetes, respectively 30,31 . Also, ARL6IP1 protein is associated phosphatidylethanolamine-binding family of proteins (PEBP1), which has important role in MAPK, NF-kappa B, and glycogen synthase kinase-3 (GSK-3) signaling pathways.…”

Section: Discussionmentioning

confidence: 99%

Altered pathways in methylome and transcriptome longitudinal analysis of normal weight and bariatric surgery women

Nicoletti

Pinhel

Noronha

et al. 2020

Sci Rep

View full text Add to dashboard Cite

DnA methylation could provide a link between environmental, genetic factors and weight control and can modify gene expression pattern. This study aimed to identify genes, which are differentially expressed and methylated depending on adiposity state by evaluating normal weight women and obese women before and after bariatric surgery (BS). We enrolled 24 normal weight (BMI: 22.5 ± 1.6 kg/m 2 ) and 24 obese women (BMI: 43.3 ± 5.7 kg/m 2 ) submitted to BS. Genome-wide methylation analysis was conducted using Infinium Human Methylation 450 BeadChip (threshold for significant CpG sites based on delta methylation level with a minimum value of 5%, a false discovery rate correction (FDR) of q < 0.05 was applied). Expression levels were measured using HumanHT-12v4 Expression BeadChip (cutoff of p ≤ 0.05 and fold change ≥2.0 was used to detect differentially expressed probes). The integrative analysis of both array data identified four genes (i.e. TPP2, PSMG6, ARL6IP1 and FAM49B) with higher methylation and lower expression level in pre-surgery women compared to normal weight women: and two genes (i.e. ZFP36L1 and USP32) that were differentially methylated after BS. These methylation changes were in promoter region and gene body. All genes are related to MAPK cascade, NIK/NF-kappaB signaling, cellular response to insulin stimulus, proteolysis and others. integrating analysis of DnA methylation and gene expression evidenced that there is a set of genes relevant to obesity that changed after BS. A gene ontology analysis showed that these genes were enriched in biological functions related to adipogenesis, orexigenic, oxidative stress and insulin metabolism pathways. Also, our results suggest that although methylation plays a role in gene silencing, the majority of effects were not correlated.DNA methylation in CpG dinucleotides is a dynamic process and best characterized epigenetic modifications 1 . Methylation patterns are established in early life and can be remodeled in adult cells, by modulating DNA interactions with proteins and transcription factors 2,3 , being able to alter gene expression profile 4 .Weight gain or loss during infancy and adulthood, by changing the energy storage and adipose tissue homeostasis, may alter molecular mechanisms and biological processes 5 . In line of this, epigenetic changes may predispose a disease risk or can occur once a disease has developed 6 . Thus, DNA methylation provides a link between environmental, genetic factors and weight control.Obesity has reached epidemic proportions and, in 2016, affected about 1.9 billion adults worldwide 7 , being associated to a public health burden. In the context of obesity treatment, bariatric surgery (BS) has been the best choice for cases of severe obesity and has been shown to be the most effective way to promote significant and sustained weight loss 8 . Among the surgical techniques, Roux-en-Y gastric bypass (RYGB) is the most performed in Brazil and worldwide, and is considered a gold standard because its high efficacy and low morbid-mortalit...

show abstract

Determination of topological structure of ARL6ip1 in cells: Identification of the essential binding region of ARL6ip1 for conophylline

Cited by 15 publications

References 17 publications

Topological analysis of DPY19L3, a human C‐mannosyltransferase

Topological analysis of DPY19L3, a human C‐mannosyltransferase

Pentylenetetrazol modulates redox system by inducing addicsin translocation from endoplasmic reticulum to plasma membrane in NG108-15 cells

Altered pathways in methylome and transcriptome longitudinal analysis of normal weight and bariatric surgery women

Contact Info

Product

Resources

About