A mitochondrial FUNDC1/HSC70 interaction organizes the proteostatic stress response at the risk of cell morbidity

Li, Yanjun; Xue, Yanhong; Xu, Xiao‐Jun; Wang, Guopeng; Liu, Yiqun; Wu, Hao; Li, Wenhui; Wang, Yueying; Chen, Ziheng; Zhang, Weilin; Zhu, Yushan; Ji, Wei; Xu, Tao; Liu, Lei; Chen, Quan

doi:10.15252/embj.201798786

Cited by 91 publications

(80 citation statements)

References 54 publications

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“…has been shown to be involved in delivery of cytosolic proteins to the nucleus, mitochondrion and endoplasmic reticulum. 24,25 Due to this interaction and the observation that Hsc70 recognizes mitochondrial aspartate aminotransferase but not cytosolic, 26,27 we reason that our protocol is consistent with the expectation of producing biologically relevant interactions. [28][29][30][31] Based on these data we proceeded to analyze all 115 visible bands between 10 and 250 kDa in Figure 1A.…”

Section: Resultssupporting

confidence: 62%

Lipopolysaccharide stimulation of RAW264.7 cells is a model for identifying novel clients of Hsc70

Rakus

et al. 2020

Preprint

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Heat shock cognate protein 71 kDa (Hsc70, Hspa8, Hspa10, Hsp73) is a member of the heat shock protein 70 kDa family of molecular chaperones. These chaperones function to aid the correct folding of client proteins using an ATP-dependent mechanism. Though Hsc70 is accepted to be a constitutively expressed protein, it has a well-documented function in modulating the induction of the pro-inflammatory cytokine TNFα by the LPS/TLR4 pathway. In this work we attempt to identify protein clients of Hsc70 to gain insight into those which may be responsible for this regulatory effect. RAW264.7 cells were cultured in the absence or presence of 1 μ g/mL LPS for 0 to 24 hours. Herein we describe of a large number of newly-categorized Hsc70 clients using immunoprecipitation and nanoLC-MS/MS and we validate several novel Hsc70/client interactions using co-immunoprecipitation. After performing immunoprecipitation using a commercially available antibody, eluted fractions were proteolytically digested either immediately after immunoprecipitation or after separation by SDS-PAGE and sequence analyzed using nanoLC-MS/MS with a Q-Exactive Plus triple-quadrupole/orbitrap mass spectrometer.Using these methods, 292 total unique protein hits were identified with high confidence; 34 of which were only detected in LPS-treated cells.

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Section: Resultssupporting

confidence: 62%

Lipopolysaccharide stimulation of RAW264.7 cells is a model for identifying novel clients of Hsc70

Rakus

et al. 2020

Preprint

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“…The stressors leading to senescence include telomere shortening due to replicative exhaustion, DNA or chromatin structure and mitochondrial dysfunction [15][16][17][18][19]. Recent reports indicated that mitochondrial dysfunction plays an important role in initiating inflammatory responses and cellular senescence and has significance in the pathogenesis of lung disease [20]. ROS generation occurs via tissue damageactivated RAC1 and dysfunctional mitochondrial.…”

Section: Discussionmentioning

confidence: 99%

miR-200b regulates cellular senescence and inflammatory responses by targeting ZEB2 in pulmonary emphysema

Shen

Xuan

Wang

et al. 2020

Artificial Cells, Nanomedicine, and Biotechnology

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Smoking is an important factor in the pathogenesis of chronic obstructive pulmonary disease (COPD), which is commonly characterised by cellular senescence and inflammation. Recently, miR-200b has emerged as an important target to cure lung disease; however, the function of miR-200b in reducing cellular senescence and inflammatory responses has not been reported. In this study, we found that miR-200b was downregulated in the lungs of COPD model mice, and its expression is correlated with cellular senescence and inflammatory responses. We hypothesised that miR-200b may be a potential novel therapy for treating COPD. We performed senescence-Associated-b-galactosidase (SA-b-GAL) staining, western blot, qRT-PCR and ELISA; our data suggested that miR-200b is an anti-aging factor in the lungs that is involved in inflammatory responses. We also confirmed that ZEB2 (Zinc finger E-box binding homeobox 2) is a target gene of miR-200b using luciferase reporter assay. In addition, we verified the function of ZEB2 in cellular senescence and inflammatory responses through transfection experiments. Moreover, we found that the protective effects of miR-200b are inhibited when cells overexpress the ZEB2 protein. In conclusion, our results suggest that miR-200b may attenuate cellular senescence and inflammatory responses by targeting ZEB2 in pulmonary emphysema.

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“…The second group of mitophagy receptors [20] contains a transmembrane domain that can directly bind to the OMM and stimulate autophagosome formation via their LIR motifs, namely Bnip3 (BCL2 interacting protein 3) [71], Nix/Bnip3L (BCL2-like 3) [72], FUNDC1 (FUN14 domain containing 1) [73] and Bcl2L13 (BCL2-like 13) [74]. Their roles in the regulation of cell proliferation, cytosolic protein degradation and cell death have also been reported [75][76][77]. However, their importance in mitochondrial clearance is not clearly known.…”

Section: Conflicts Of Interestmentioning

confidence: 99%

Mitophagy in the Retinal Pigment Epithelium of Dry Age-Related Macular Degeneration Investigated in the NFE2L2/PGC-1α-/- Mouse Model

Gurubaran

Viiri

Koskela

et al. 2020

IJMS

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Increased oxidative stress and mitochondrial damage are observed in protein aggregation diseases, such as age-related macular degeneration (AMD). We have recently reported elevated levels of oxidative stress markers, damaged mitochondria, accumulating lysosomal lipofuscin and extracellular drusen-like structures in the retinal pigment epithelial cells (RPE) of the dry AMD-resembling NFE2L2/PGC1α double knockout (dKO) mouse model. Here, we provide evidence of a disturbance in the autolysosomal machinery handling mitochondrial clearance in the RPE cells of one-year-old NFE2L2/PGC1α-deficient mice. Confocal immunohistochemical analysis revealed an upregulation of autophagosome marker microtubule-associated proteins 1A/1B light chain 3B (LC3B) as well as numerous mitophagy markers, such as PTE-induced putative kinase 1 (PINK1) and E3 ubiquitin ligase (PARKIN) together with damaged mitochondria. However, we detected no evidence of increased autolysosome formation in transmission electron micrographs or of colocalization of lysosomal marker LAMP2 (lysosome-associated membrane protein 2) and the mitochondrial marker ATP synthase β in confocal micrographs. Interestingly, we observed an upregulation of late autolysosomal fusion Ras-related protein (Rab7) in the perinuclear space of RPE cells together with autofluorescence aggregates. Our results reveal that there is at least a relative decrease of mitophagy in the RPE cells of NFE2L2/PGC1α dKO mice. This further supports the hypothesis that mitophagy is a putative therapy target in AMD-like pathology.

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A mitochondrial FUNDC1/HSC70 interaction organizes the proteostatic stress response at the risk of cell morbidity

Cited by 91 publications

References 54 publications

Lipopolysaccharide stimulation of RAW264.7 cells is a model for identifying novel clients of Hsc70

Lipopolysaccharide stimulation of RAW264.7 cells is a model for identifying novel clients of Hsc70

miR-200b regulates cellular senescence and inflammatory responses by targeting ZEB2 in pulmonary emphysema

Mitophagy in the Retinal Pigment Epithelium of Dry Age-Related Macular Degeneration Investigated in the NFE2L2/PGC-1α-/- Mouse Model

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