The protective role of protein L-isoaspartyl (D-aspartate)<i>O</i>-methyltransferase for maintenance of mitochondrial morphology in A549 cell

Ogasawara, Masahito; Otani, Mieko; Takano, Masaoki; Shudou, Masachika; Inaba, Yohei; Nirasawa, Satoru; Takahashi, Saori; Kiyoi, Takeshi; Tanaka, Yuki; Kameda, Kenji; Kunugita, Naoki; Maeyama, Kazutaka; Sano, Keiji; Yamashita, Masahiro; Yamauchi, Kohei

doi:10.1080/01902148.2016.1197984

Cited by 5 publications

(6 citation statements)

References 54 publications

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“…However, whereas mitophagy is thought to affect the mitochondrion as a whole, we only found effects on certain proteins. Specifically, loss of parkin in our study correlated with alterations in the abundance of synaptic mitochondrial proteins related to protein aggregate clearance and repair mechanisms including Hsph1 , Hspa4l 88 , and Pcmt1 91 . Importantly, these have been shown to act on alpha-synuclein 88 , the major proteopathic component of Lewy Bodies which contributes to the neuropathological damage in PD 37 , 92 .…”

Section: Discussionmentioning

confidence: 51%

Longitudinal in vivo metabolic labeling reveals tissue-specific mitochondrial proteome turnover rates and proteins selectively altered by parkin deficiency

Stauch

Totusek

Trease

et al. 2023

Sci Rep

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Our study utilizes a longitudinal isotopic metabolic labeling approach in vivo in combination with organelle fraction proteomics to address the role of parkin in mitochondrial protein turnover in mice. The use of metabolic labeling provides a method to quantitatively determine the global changes in protein half-lives whilst simultaneously assessing protein expression. Studying two diverse mitochondrial populations, we demonstrated the median half-life of brain striatal synaptic mitochondrial proteins is significantly greater than that of hepatic mitochondrial proteins (25.7 vs. 3.5 days). Furthermore, loss of parkin resulted in an overall, albeit modest, increase in both mitochondrial protein abundance and half-life. Pathway and functional analysis of our proteomics data identified both known and novel pathways affected by loss of parkin that are consistent with its role in both mitochondrial quality control and neurodegeneration. Our study therefore adds to a growing body of evidence suggesting dependence on parkin is low for basal mitophagy in vivo and provides a foundation for the investigation of novel parkin targets.

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Section: Discussionmentioning

confidence: 51%

Longitudinal in vivo metabolic labeling reveals tissue-specific mitochondrial proteome turnover rates and proteins selectively altered by parkin deficiency

Stauch

Totusek

Trease

et al. 2023

Sci Rep

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“…Cells were grown in RPMI1640 media supplemented with 10% (v/v) fetal bovine serum (FBS), penicillin, and streptomycin, in a humidified atmosphere of 5% CO 2 and 95% air at 37° C. The cells were tested and authenticated by short tandem repeat analysis. Stably transformed cells were selected using geneticin (1000 μg/mL) and collected by cell sorting three times (>95% purified) with a fluorescent protein marker (ZsGreen1) to improve cell purity [ 41 ].…”

Section: Methodsmentioning

confidence: 99%

“…A PIMT mRNA expression vector was generated by alternative splicing [ 41 ], and a pIRES2-ZsGreen1 vector was used to express PIMT. PCMT1-ER cDNA was amplified by PCR using the forward primer 5′-CG GAA TTC ATG CCG GGA GCG CGC AGT GGC GGC AGC-3′ (underlined sequence is an EcoRI cleavage site) and reverse primer 5′-GC GGA TCC TTA CAA TTC ATC CCT GGA CCA CTG C-3′ (underlined sequence is a BamHI cleavage site).…”

Section: Methodsmentioning

confidence: 99%

Deficiency of protein-L-isoaspartate (D-aspartate) O-methyl-transferase expression under endoplasmic reticulum stress promotes epithelial mesenchymal transition in lung adenocarcinoma

et al. 2018

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A prognostic association between the novel chaperone protein-L-isoaspartate (D-aspartate) O-methyltransferase (PIMT) and lung adenocarcinoma has recently been reported. Here, we evaluated the functional roles of PIMT in the progression of lung adenocarcinoma. PIMT expression was detectable in 6 lung adenocarcinoma cell lines: A549, H441, H460, H1650, Calu 1, and Calu 6 cell lines. In A549 and H441 cells, knockdown by PIMT using silencing RNA of PIMT (si-PIMT) and/or small hairpin-RNA (sh-PIMT) induced a decrease in the expression of E-cadherin with an increase in vimentin expression, indicating that the epithelial to mesenchymal transition (EMT) was induced. Cell mobility, including migration and invasion capability, was increased in sh-PIMT A549 stable and si-PIMT H441 cells compared to in control cells. Endoplasmic reticulum (ER) stress, such as Thapsigargin (Tg) stress and hypoxia, induced EMT in A549 cells but not in other cell types, with an increase in GRP78 expression, whereas overexpression of PIMT reduced the EMT and cell invasion under stress conditions. The expression of hypoxia inducible factor-1 alpha (HIF1α) and Twist increased in sh-PIMT A549 and si-PIMT H441 cells, and Tg stress increased HIF1α expression levels in A549 cells in a dose-dependent manner. Moreover, LW6, an HIF1α inhibitor, reduced EMT, cancer invasion, and the levels of Twist in sh-PIMT A549 cells. Our results indicate that deficiency of supplemental PIMT expression under ER stress facilitates EMT and cell invasion in some cell types of lung adenocarcinoma.

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“…Impaired release or re-uptake of calcium from subcellular organelles such as the ER or the mitochondria may also contribute to the observed calcium phenotypes. Accordingly, PCMT1-deficient A549 cells were recently reported to display mitochondrial dysmorphology and decreased intracellular ATP levels (Ogasawara et al, 2016). Compromised energy levels may have led to the virtually absent calcium response to the second ATP stimulation in the HT22 Pcmt1 knockout cells.…”

Section: Interplay Between Pcmt and Calciummentioning

confidence: 99%

l-Isoaspartyl Methyltransferase Deficiency in Zebrafish Leads to Impaired Calcium Signaling in the Brain

et al. 2021

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Isomerization of l-aspartyl and l-asparaginyl residues to l-isoaspartyl residues is one type of protein damage that can occur under physiological conditions and leads to conformational changes, loss of function, and enhanced protein degradation. Protein l-isoaspartyl methyltransferase (PCMT) is a repair enzyme whose action initiates the reconversion of abnormal l-isoaspartyl residues to normal l-aspartyl residues in proteins. Many lines of evidence support a crucial role for PCMT in the brain, but the mechanisms involved remain poorly understood. Here, we investigated PCMT activity and function in zebrafish, a vertebrate model that is particularly well-suited to analyze brain function using a variety of techniques. We characterized the expression products of the zebrafish PCMT homologous genes pcmt and pcmtl. Both zebrafish proteins showed a robust l-isoaspartyl methyltransferase activity and highest mRNA transcript levels were found in brain and testes. Zebrafish morphant larvae with a knockdown in both the pcmt and pcmtl genes showed pronounced morphological abnormalities, decreased survival, and increased isoaspartyl levels. Interestingly, we identified a profound perturbation of brain calcium homeostasis in these morphants. An abnormal calcium response upon ATP stimulation was also observed in mouse hippocampal HT22 cells knocked out for Pcmt1. This work shows that zebrafish is a promising model to unravel further facets of PCMT function and demonstrates, for the first time in vivo, that PCMT plays a pivotal role in the regulation of calcium fluxes.

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The protective role of protein L-isoaspartyl (D-aspartate)O-methyltransferase for maintenance of mitochondrial morphology in A549 cell

Abstract: The increased proportion of the D-Asp residues may contribute to COPD pathogenesis, via irreversible protein conformational changes, followed by mitochondrial dysfunction.

Cited by 5 publications

References 54 publications

Longitudinal in vivo metabolic labeling reveals tissue-specific mitochondrial proteome turnover rates and proteins selectively altered by parkin deficiency

Longitudinal in vivo metabolic labeling reveals tissue-specific mitochondrial proteome turnover rates and proteins selectively altered by parkin deficiency

Deficiency of protein-L-isoaspartate (D-aspartate) O-methyl-transferase expression under endoplasmic reticulum stress promotes epithelial mesenchymal transition in lung adenocarcinoma

l-Isoaspartyl Methyltransferase Deficiency in Zebrafish Leads to Impaired Calcium Signaling in the Brain

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