Hsp70 chaperone rescues C6 rat glioblastoma cells from oxidative stress by sequestration of aggregating GAPDH

Lazarev, Vladimir F.; Nikotina, Alina D.; Mikhaylova, Elena R.; Nudler, Evgeny; Полоник, С. Г.; Guzhova, Irina V.; Margulis, Boris A.

doi:10.1016/j.bbrc.2015.12.076

Cited by 31 publications

(19 citation statements)

References 25 publications

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“…Callahan et al demonstrated that oxidative conditions lead to conformational changes which enhance the peptide accessibility to the peptide-binding-pocket. Lazarev et al found that Hsp70 binds to oxidized GAPDH, preventing it from further damage in C6 rat glioblastoma cells [54]. We were able to confirm these data by demonstrating the ability of Hsp70 to bind oxidized proteins in vitro, as well as in our cell model and in vivo (Fig.…”

Section: Discussionsupporting

confidence: 84%

The molecular chaperone Hsp70 promotes the proteolytic removal of oxidatively damaged proteins by the proteasome

Reeg

Jung

Castro

et al. 2016

Free Radical Biology and Medicine

102

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One hallmark of aging is the accumulation of protein aggregates, promoted by the unfolding of oxidized proteins. Unraveling the mechanism by which oxidized proteins are degraded may provide a basis to delay the early onset of features, such as protein aggregate formation, that contribute to the aging phenotype. In order to prevent aggregation of oxidized proteins, cells recur to the 20S proteasome, an efficient turnover proteolysis complex. It has previously been shown that upon oxidative stress the 26S proteasome, another form, dissociates into the 20S form. A critical player implicated in its dissociation is the Heat Shock Protein 70 (Hsp70), which promotes an increase in free 20S proteasome and, therefore, an increased capability to degrade oxidized proteins. The aim of this study was to test whether or not Hsp70 is involved in cooperating with the 20S proteasome for a selective degradation of oxidatively damaged proteins. Our results demonstrate that Hsp70 expression is induced in HT22 cells as a result of mild oxidative stress conditions. Furthermore, Hsp70 prevents the accumulation of oxidized proteins and directly promotes their degradation by the 20S proteasome. In contrast the expression of the Heat shock cognate protein 70 (Hsc70) was not changed in recovery after oxidative stress and Hsc70 has no influence on the removal of oxidatively damaged proteins. We were able to demonstrate in HT22 cells, in brain homogenates from 129/SV mice and in vitro, that there is an increased interaction of Hsp70 with oxidized proteins, but also with the 20S proteasome, indicating a role of Hsp70 in mediating the interaction of oxidized proteins with the 20S proteasome. Thus, our data clearly implicate an involvement of Hsp70 oxidatively damaged protein degradation by the 20S proteasome.

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Section: Discussionsupporting

confidence: 84%

The molecular chaperone Hsp70 promotes the proteolytic removal of oxidatively damaged proteins by the proteasome

Reeg

Jung

Castro

et al. 2016

Free Radical Biology and Medicine

102

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“…Several studies have shown that HSP70, a BIS binding partner, has a protective function as a chaperone protein under oxidative stress conditions. 27 , 28 Given the decrease in cell viability induced by H 2 O 2 upon deletion of BIS residues 281–292, we hypothesized that HSP70 along with BIS protects against H 2 O 2 . HSP70 levels were unaffected by H 2 O 2 treatment in cells overexpressing the empty vector or FL-BIS; however, in Δ281–292 BIS-transfected cells, HSP70 expression was reduced under oxidative stress conditions.…”

Section: Resultsmentioning

confidence: 99%

ERK-mediated phosphorylation of BIS regulates nuclear translocation of HSF1 under oxidative stress

Kim

Yun

et al. 2016

Exp Mol Med

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B-cell lymphoma (BCL)-2-interacting cell death suppressor (BIS) has diverse cellular functions depending on its binding partners. However, little is known about the effects of biochemical modification of BIS on its various activities under oxidative stress conditions. In this study, we showed that H2O2 reduced BIS mobility on SDS–polyacrylamide gels in a time-dependent manner via the activation of extracellular signaling-regulated kinase (ERK). The combined results of mass spectroscopy and computational prediction identified Thr285 and Ser289 in BIS as candidate residues for phosphorylation by ERK under oxidative stress conditions. Deletion of these sites resulted in a partial reduction in the H2O2-induced mobility shift relative to that of the wild-type BIS protein; overexpression of the deletion mutant sensitized A172 cells to H2O2-induced cell death without increasing the level of intracellular reactive oxygen species. Expression of the BIS deletion mutant decreased the level of heat shock protein (HSP) 70 mRNA following H2O2 treatment, which was accompanied by impaired nuclear translocation of heat shock transcription factor (HSF) 1. Co-immunoprecipitation assays revealed that the binding of wild-type BIS to HSF1 was decreased by oxidative stress, while the binding of the BIS deletion mutant to HSF1 was not affected. These results indicate that ERK-dependent phosphorylation of BIS has a role in the regulation of nuclear translocation of HSF1 likely through modulation of its interaction affinity with HSF1, which affects HSP70 expression and sensitivity to oxidative stress.

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“…Therefore, we used 3 GAPDH fractions: to verify the aggregate content in each fraction we applied ultrafiltration through acetate cellulose membrane ( Fig. 1 ) as described earlier [4] . To explore the dynamics of apoptosis, the cells were placed in a 96-well micro-titre plate, and acridine orange in phosphate-buffered saline was added to a concentration of 5 μg/mL.…”

Section: Experimental Design Materials and Methodsmentioning

confidence: 99%

Hydrocortisone 21-hemisuccinate did not prevent exogenous GAPDH-induced apoptosis in human neuroblastoma cells

et al. 2018

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These data are related to our paper “GAPDH-targeted therapy – a new approach for secondary damage after traumatic brain injury on rats” (Lazarev et al., In press), in which we explore the role of exogenous GAPDH in traumatic brain injury-induced neuron death, and the therapeutic application of small molecules that bind to the enzyme. The current article demonstrates the induction of apoptosis by exogenous GAPDH and the effectiveness of the hydrocortisone derivative for suppressing the pathogenic action of the enzyme.

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Hsp70 chaperone rescues C6 rat glioblastoma cells from oxidative stress by sequestration of aggregating GAPDH

Cited by 31 publications

References 25 publications

The molecular chaperone Hsp70 promotes the proteolytic removal of oxidatively damaged proteins by the proteasome

The molecular chaperone Hsp70 promotes the proteolytic removal of oxidatively damaged proteins by the proteasome

ERK-mediated phosphorylation of BIS regulates nuclear translocation of HSF1 under oxidative stress

Hydrocortisone 21-hemisuccinate did not prevent exogenous GAPDH-induced apoptosis in human neuroblastoma cells

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