La Autoantigen Mediates Oxidant Induced De Novo Nrf2 Protein Translation

Zhang, Jack; Dinh, Thai Nho; Kappeler, Kyle V.; Tsaprailis, George; Chen, Qin M.

doi:10.1074/mcp.m111.015032

Cited by 43 publications

(66 citation statements)

References 43 publications

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“…These data indicate that both cytochrome b and cytochrome c expression decreased due to H 2 O 2 treatment. (10,18,19). To address whether this condition of inducing Nrf2 protein results in protection against mitochondrial damage, cells were pretreated with 100 mM H 2 O 2 for 10 min, followed by a 1 h recovery period.…”

Section: Discussionmentioning

confidence: 99%

Nrf2 protects mitochondrial decay by oxidative stress

et al. 2015

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Sublethal levels of oxidative stress are commonly associated with various pathophysiological conditions. Cardiomyocytes have the highest content of mitochondria among all cell types, allowing the study of mitochondria in cells surviving oxidative stress and address whether nuclear factor-erythroid-derived 2-related factor 2 (Nrf2) can reverse these changes. Mitochondria normally exist in elaborated networks, which were replaced by predominately individual punctuate mitochondria 24 h after exposure to a nonlethal dose of H 2 O 2 . Electron microscopy revealed that cells surviving H 2 O 2 show swelling of mitochondria with disorganized cristae and areas of condensation. Measurements of functional mitochondria showed a H 2 O 2 dosedependent decrease over a course of 5 d. At the protein and mRNA levels, cells surviving H 2 O 2 treatment show a reduction of mitochondrial components, cytochrome c, and cytochrome b. Nrf2 overexpression prevented H 2 O 2 from inducing mitochondria morphologic changes and reduction of cytochrome b/c. Although Nrf2 is known as a transcription factor regulating antioxidant and detoxification genes, Nrf2 overexpression did not significantly reduce the level of protein oxidation. Instead, Nrf2 was found to associate with the outer mitochondrial membrane. Mitochondria prepared from the myocardium of Nrf2 knockout mice are more sensitive to permeability transition. Our data suggest that Nrf2 protects mitochondria from oxidant injury likely through direct interaction with mitochondria.-Strom, J., Xu, B., Tian, X., Chen, Q. M. Nrf2 protects mitochondrial decay by oxidative stress. FASEB J. 30, 66-80 (2016). www.fasebj.org

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Section: Discussionmentioning

confidence: 99%

Nrf2 protects mitochondrial decay by oxidative stress

et al. 2015

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“…13A). This differs from La autoantigen, an RNA binding protein capable of binding to NRF2 mRNA and increasing the association with 60/80S ribosomal fractions during oxidative stress (17). The association of EF1a with ribosomal small or large subunits did not appear to change due to H 2 O 2 treatment, as measured by coimmunoprecipitation of the large-or small-subunit protein L36a or S6, respectively (Fig.…”

Section: Figmentioning

confidence: 95%

“…The C-terminal half of the NRF2 protein contains a cap-and-collar domain, a basic amino acid region for DNA binding, and a leucine zipper for heterodimerization with a cotranscription factor, such as the small musculoaponeurotic factor (12)(13)(14)(15)(16). Although the NRF2 protein can be stabilized due to chemically induced dissociation from KEAP1, we have found that oxidative stress induces de novo NRF2 protein translation in vitro and in vivo (7,8,17).…”

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confidence: 91%

“…For the A4 mutant sequence, the GGGG sequence in the region spanning nt Ϫ195 to Ϫ191 was replaced with TTTT in the template. RNA affinity chromatography was performed to pull down proteins, as described previously (17,88). Briefly, HeLa cells were harvested in nucleic acid binding (NAB) buffer (10 mM HEPES [pH 7.6], 5 mM MgCl 2 , 40 mM KCl, 1 mM dithiothreitol, 5% glycerol, 5 mg/ml heparin, 1 U/ml RNasin), followed by 3 brief sonications (5 s each) on ice.…”

Section: Methodsmentioning

confidence: 99%

“…The RNA binding proteins were released by boiling in SDS-PAGE loading buffer, separated by SDS-PAGE, and identified by mass spectrometry-compatible silver staining (Bio-Rad). Protein bands showing differential binding between the control and H 2 O 2 treatments were excised for LC-MS/MS analyses as described previously (17).…”

Section: Methodsmentioning

confidence: 99%

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G-Quadruplex in the NRF2 mRNA 5′ Untranslated Region Regulates De Novo NRF2 Protein Translation under Oxidative Stress

Lee

Zhang

Strom

et al. 2017

Molecular and Cellular Biology

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Inhibition of protein synthesis serves as a general measure of cellular consequences of chemical stress. A few proteins are translated selectively and influence cell fate. How these proteins can bypass the general control of translation remains unknown. We found that low to mild doses of oxidants induce de novo translation of the NRF2 protein. Here we demonstrate the presence of a G-quadruplex structure in the 5= untranslated region (UTR) of NRF2 mRNA, as measured by circular dichroism, nuclear magnetic resonance, and dimethylsulfate footprinting analyses. Such a structure is important for 5=-UTR activity, since its removal by sequence mutation eliminated H 2 O 2 -induced activation of the NRF2 5= UTR. Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based proteomics revealed elongation factor 1 alpha (EF1a) as a protein binding to the G-quadruplex sequence. Cells responded to H 2 O 2 treatment by increasing the EF1a protein association with NRF2 mRNA, as measured by RNA-protein interaction assays. The EF1a interaction with small and large subunits of ribosomes did not appear to change due to H 2 O 2 treatment, nor did posttranslational modifications, as measured by two-dimensional (2-D) Western blot analysis. Since NRF2 encodes a transcription factor essential for protection against tissue injury, our data have revealed a novel mechanism of cellular defense involving de novo NRF2 protein translation governed by the EF1a interaction with the G-quadruplex in the NRF2 5= UTR during oxidative stress.

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The RNA‐binding protein La/SSB associates with radiation‐induced DNA double‐strand breaks in lung cancer cell lines

Staudacher

Liapis

et al. 2021

Cancer Reports

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Background Platinum‐based chemotherapy and radiotherapy are standard treatments for non‐small cell lung cancer, which is the commonest, most lethal cancer worldwide. As a marker of treatment‐induced cancer cell death, we have developed a radiodiagnostic imaging antibody, which binds to La/SSB. La/SSB is an essential, ubiquitous ribonuclear protein, which is over expressed in cancer and plays a role in resistance to cancer therapies. Aim In this study, we examined radiation‐induced DNA double strand breaks (DSB) in lung cancer cell lines and examined whether La/SSB associated with these DSB. Method Three lung cancer lines (A549, H460 and LL2) were irradiated with different X‐ray doses or X‐radiated with a 5 Gy dose and examined at different time‐points post‐irradiation for DNA DSB in the form of γ‐H2AX and Rad51 foci. Using fluorescence microscopy, we examined whether La/SSB and γ‐H2AX co‐localise and performed proximity ligation assay (PLA) and co‐immunoprecipitation to confirm the interaction of these proteins. Results We found that the radio‐resistant A549 cell line compared to the radio‐sensitive H460 cell line showed faster resolution of radiation‐induced γ‐H2AX foci over time. Conversely, we found more co‐localised γ‐H2AX and La/SSB foci by PLA in irradiated A549 cells. Conclusion The co‐localisation of La/SSB with radiation‐induced DNA breaks suggests a role of La/SSB in DNA repair, however further experimentation is required to validate this.

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La Autoantigen Mediates Oxidant Induced De Novo Nrf2 Protein Translation

Cited by 43 publications

References 43 publications

Nrf2 protects mitochondrial decay by oxidative stress

Nrf2 protects mitochondrial decay by oxidative stress

G-Quadruplex in the NRF2 mRNA 5′ Untranslated Region Regulates De Novo NRF2 Protein Translation under Oxidative Stress

The RNA‐binding protein La/SSB associates with radiation‐induced DNA double‐strand breaks in lung cancer cell lines

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