Regulation of plant ER oxidoreductin 1 (ERO1) activity for efficient oxidative protein folding

Matsusaki, Motonori; Okuda, Aya; Matsuo, Koichi; Gekko, Kunihiko; Masuda, Taro; Naruo, Yurika; Hirose, Akiho; Kono, Kentaro; Tsuchi, Yuichiro; Urade, Reiko

doi:10.1074/jbc.ra119.010917

Cited by 19 publications

(17 citation statements)

References 63 publications

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“…It has been reported that ERO1L is transcriptionally regulated by the ER unfolded protein response 25 , 26 . To distinguish this possibility in PDAC, we assessed whether ERO1L expression is induced upon treatment with three established chemical inducers of ER stress: thapsigargin, tunicamycin, and dithiothreitol (DTT).…”

Section: Resultsmentioning

confidence: 99%

Endoplasmic Reticulum stress-dependent expression of ERO1L promotes aerobic glycolysis in Pancreatic Cancer

Zhang¹,

Yang²,

Lin³

et al. 2020

Theranostics

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Rationale: Endoplasmic reticulum oxidoreductase 1 alpha (ERO1L) is an endoplasmic reticulum (ER) luminal glycoprotein that has a role in the formation of disulfide bonds of secreted proteins and membrane proteins. Emerging data identify ERO1L as a tumor promoter in a wide spectrum of human malignancies. However, its molecular basis of oncogenic activities remains largely unknown. Methods: Pan-cancer analysis was performed to determine the expression profile and prognostic value of ERO1L in human cancers. The mechanism by which ERO1L promotes tumor growth and glycolysis in pancreatic ductal adenocarcinoma (PDAC) was investigated by cell biological, molecular, and biochemical approaches. Results: ERO1L was highly expressed in PDAC and its precursor pancreatic intraepithelial neoplasia and acts as an independent prognostic factor for patient survival. Hypoxia and ER stress contributed to the overexpression pattern of ERO1L in PDAC. ERO1L knockdown or pharmacological inhibition with EN460 suppressed PDAC cell proliferation in vitro and slowed tumor growth in vivo . Ectopic expression of wild type ERO1L but not its inactive mutant form EROL-C394A promoted tumor growth. Bioinformatics analyses and functional analyses confirmed a regulatory role of ERO1L on the Warburg effect. Notably, inhibition of tumor glycolysis partially abrogated the growth-promoting activity of ERO1L. Mechanistically, ERO1L-mediated ROS generation was essential for its oncogenic activities. In clinical samples, ERO1L expression was correlated with the maximum standard uptake value (SUVmax) in PDAC patients who received 18 F-FDG PET/CT imaging preoperatively. Analysis of TCGA cohort revealed a specific glycolysis gene expression signature that is highly correlated with unfolded protein response-related gene signature. Conclusion: Our findings uncover a key function for ERO1L in Warburg metabolism and indicate that targeting this pathway may offer alternative therapeutic strategies for PDAC.

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Section: Resultsmentioning

confidence: 99%

Endoplasmic Reticulum stress-dependent expression of ERO1L promotes aerobic glycolysis in Pancreatic Cancer

Zhang¹,

Yang²,

Lin³

et al. 2020

Theranostics

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“…To investigate complex formation between P5 and other PDIs via non-covalent interactions, we used the far-western dot blot method [ 44 , 45 , 55 , 56 ] ( Figure 1 b,c). Briefly, we fixed known concentrations of reduced forms of PDIs as prey proteins onto a nitrocellulose membrane.…”

Section: Resultsmentioning

confidence: 99%

“…Far-western dot blot analyses were performed by optimizing the method reported in previous studies [ 44 , 45 ]. Reduced purified protein solutions (40 µM) were diluted to an appropriate concentration, and a 3 µL aliquot was dotted onto a ClearTrans nitrocellulose membrane (Cat.…”

Section: Methodsmentioning

confidence: 99%

Functional Interplay between P5 and PDI/ERp72 to Drive Protein Folding

Matsusaki

Okada

Tanikawa

et al. 2021

Biology

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P5 is one of protein disulfide isomerase family proteins (PDIs) involved in endoplasmic reticulum (ER) protein quality control that assists oxidative folding, inhibits protein aggregation, and regulates the unfolded protein response. P5 reportedly interacts with other PDIs via intermolecular disulfide bonds in cultured cells, but it remains unclear whether complex formation between P5 and other PDIs is involved in regulating enzymatic and chaperone functions. Herein, we established the far-western blot method to detect non-covalent interactions between P5 and other PDIs and found that PDI and ERp72 are partner proteins of P5. The enzymatic activity of P5-mediated oxidative folding is up-regulated by PDI, while the chaperone activity of P5 is stimulated by ERp72. These findings shed light on the mechanism by which the complex formations among PDIs drive to synergistically accelerate protein folding and prevents aggregation. This knowledge has implications for understanding misfolding-related pathology.

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“…The method has been used for the structural analysis of unknown proteins in the native and other states so far ( 22 , 23 ).…”

Section: Methodsmentioning

confidence: 99%

Secondary Structure of Human De Novo Evolved Gene Product NCYM Analyzed by Vacuum-Ultraviolet Circular Dichroism

et al. 2021

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NCYM, a cis-antisense gene of MYCN, encodes a Homininae-specific protein that promotes the aggressiveness of human tumors. Newly evolved genes from non-genic regions are known as de novo genes, and NCYM was the first de novo gene whose oncogenic functions were validated in vivo. Targeting NCYM using drugs is a potential strategy for cancer therapy; however, the NCYM structure must be determined before drug design. In this study, we employed vacuum-ultraviolet circular dichroism to evaluate the secondary structure of NCYM. The SUMO-tagged NCYM and the isolated SUMO tag in both hydrogenated and perdeuterated forms were synthesized and purified in a cell-free in vitro system, and vacuum-ultraviolet circular dichroism spectra were measured. Significant differences between the tagged NCYM and the isolated tag were evident in the wavelength range of 190–240 nm. The circular dichroism spectral data combined with a neural network system enabled to predict the secondary structure of NCYM at the amino acid level. The 129-residue tag consists of α-helices (approximately 14%) and β-strands (approximately 29%), which corresponded to the values calculated from the atomic structure of the tag. The 238-residue tagged NCYM contained approximately 17% α-helices and 27% β-strands. The location of the secondary structure predicted using the neural network revealed that these secondary structures were enriched in the Homininae-specific region of NCYM. Deuteration of NCYM altered the secondary structure at D90 from an α-helix to another structure other than α-helix and β-strand although this change was within the experimental error range. All four nonsynonymous single-nucleotide polymorphisms (SNPs) in human populations were in this region, and the amino acid alteration in SNP N52S enhanced Myc-nick production. The D90N mutation in NCYM promoted NCYM-mediated MYCN stabilization. Our results reveal the secondary structure of NCYM and demonstrated that the Homininae-specific domain of NCYM is responsible for MYCN stabilization.

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Regulation of plant ER oxidoreductin 1 (ERO1) activity for efficient oxidative protein folding

Cited by 19 publications

References 63 publications

Endoplasmic Reticulum stress-dependent expression of ERO1L promotes aerobic glycolysis in Pancreatic Cancer

Endoplasmic Reticulum stress-dependent expression of ERO1L promotes aerobic glycolysis in Pancreatic Cancer

Functional Interplay between P5 and PDI/ERp72 to Drive Protein Folding

Secondary Structure of Human De Novo Evolved Gene Product NCYM Analyzed by Vacuum-Ultraviolet Circular Dichroism

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