10 research outputs found
Representation of Minorities in Research: A View from the Community
No full textAUTHOR AFFILIATION: Sudarshan Pyakurel, Arogya Institute and Bhutanese Nepali Community of Central Ohio, United States, [email protected] media can be accessed here:
http://streaming.osu.edu/knowledgebank/PREA/PREA_Session13C_Pyakurel_20190326.mp
Elucidation of interactors of the pro-fusion proteins MFN1 and MFN2 sheds light on novel pathways regulating mitochondrial morphology in apoptosis
No full textLarge-scale proteomics analysis provide information on the proteomic signatures including interacting partners and post-translational modification. In the first part of the thesis, we identified a novel keratin-binding protein, Trichoplein (TpMs) that requires MFN2 to modulate mitochondrial shape and tethering. In the second part, we show that the pleiotropic mitogen activated protein (MAP) kinase phosphorylate the pro-fusion protein mitofusin (MFN) 1, switching its functions in apoptosis and mitochondrial fusion. A phosphoproteomic analysis revealed that MFN1 is phosphorylated at an atypical ERK1 site (T562) of its HR1 domain. This site proved essential to mediate the MFN1-dependent mitochondrial elongation and regulation of apoptosis by the MEK/ERK pathway. A T562 mutant mimicking constitutive phosphorylation of MFN1 triggered mitochondrial fragmentation, increased susceptibility to apoptosis, bound more avidly to the proapoptotic BCL-2 family member BAK and facilitated its activation. Our data identify a role for phosphorylation of MFN1 in mitochondrial shape and apoptosis
Trichoplein/mitostatin regulates endoplasmic reticulum-mitochondria juxtaposition.
No full textTrichoplein/mitostatin (TpMs) is a keratin-binding protein that partly colocalizes with mitochondria and is often downregulated in epithelial cancers, but its function remains unclear. In this study, we report that TpMs regulates the tethering between mitochondria and endoplasmic reticulum (ER) in a Mitofusin 2 (Mfn2)-dependent manner. Subcellular fractionation and immunostaining show that TpMs is present at the interface between mitochondria and ER. The expression of TpMs leads to mitochondrial fragmentation and loosens tethering with ER, whereas its silencing has opposite effects. Functionally, the reduced tethering by TpMs inhibits apoptosis by Ca(2+)-dependent stimuli that require ER-mitochondria juxtaposition. Biochemical and genetic evidence support a model in which TpMs requires Mfn2 to modulate mitochondrial shape and tethering. Thus, TpMs is a new regulator of mitochondria-ER juxtaposition
ATR-dependent pathways control hEXO1 stability in response to stalled forks
No full textNucleases play important roles in DNA synthesis, recombination and repair. We have previously shown that human exonuclease 1 (hEXO1) is phosphorylated in response to agents stalling DNA replication and that hEXO1 consequently undergoes ubiquitination and degradation in a proteasome-dependent manner. In the present study, we have addressed the identity of the pathway transducing stalled-replication signals to hEXO1. Using chemical inhibitors, RNA interference, ATM- and ATR-deficient cell lines we have concluded that hEXO1 phosphorylation is ATR-dependent. By means of mass spectrometry, we have identified the sites of phosphorylation in hEXO1 in undamaged cells and in cells treated with hydroxyurea (HU). hEXO1 is phosphorylated at nine basal sites and three additional sites are induced by HU treatment. Analysis of single- and multiple-point mutants revealed that mutation to Ala of the three HU-induced sites of phosphorylation partially rescued HU-dependent degradation of hEXO1 and additionally stabilized the protein in non-treated cells. We have raised an antibody to pS(714), an HU-induced site of the S/T-Q type, and we provide evidence that S(714) is phosphorylated upon HU but not IR treatment. The antibody may be a useful tool to monitor signal transduction events triggered by stalled DNA replication
Bis-Retinoid A2E Induces an Increase of Basic Fibroblast Growth Factor via Inhibition of Extracellular Signal-Regulated Kinases 1/2 Pathway in Retinal Pigment Epithelium Cells and Facilitates Phagocytosis.2
No full textAge-related macular degeneration (ARMD) is the leading cause of vision loss in developed countries. Hallmarks of the disease are well known; indeed, this pathology is characterized by lipofuscin accumulation, is principally composed of lipid-containing residues of lysosomal digestion. The N-retinyl-N-retinylidene ethanolamine (A2E) retinoid which is thought to be a cytotoxic component for RPE is the best- characterized component of lipofuscin so far. Even if no direct correlation between A2E spatial distribution and lipofuscin fluorescence has been established in aged human RPE, modified forms or metabolites of A2E could be involved in ARMD pathology. Mitogen-activated protein kinase (MAPK) pathways have been involved in many pathologies, but not in ARMD. Therefore, we wanted to analyze the effects of A2E on MAPKs in polarized ARPE19 and isolated mouse RPE cells. We showed that long-term exposure of polarized ARPE19 cells to low A2E dose induces a strong decrease of the extracellular signal-regulated kinases' (ERK1/2) activity. In addition, we showed that A2E, via ERK1/2 decrease, induces a significant decrease of the retinal pigment epithelium-specific protein 65 kDa (RPE65) expression in ARPE19 cells and isolated mouse RPE. In the meantime, we showed that the decrease of ERK1/2 activity mediates an increase of basic fibroblast growth factor (bFGF) mRNA expression and secretion that induces an increase in phagocytosis via a paracrine effect. We suggest that the accumulation of deposits coming from outer segments (OS) could be explained by both an increase of bFGF-induced phagocytosis and by the decrease of clearance by A2E. The bFGF angiogenic protein may therefore be an attractive target to treat ARMD
Loss of Extracellular Signal-Regulated Kinase 1/2 in the Retinal Pigment Epithelium Leads to RPE65 Decrease and Retinal Degeneration
No full textRecent work suggested that the activity of extracellular signal-regulated kinase 1/2 (ERK1/2) is increased in the retinal pigment epithelium (RPE) of age-related macular degeneration (ARMD) patients and therefore could be an attractive therapeutic target. Notably, ERK1/2 pathway inhibitors are used in cancer therapy, with severe and noncharacterized ocular side effects. To decipher the role of ERK1/2 in RPE cells, we conditionally disrupted the Erk1 and Erk2 genes in mouse RPE. The loss of ERK1/2 activity resulted in a significant decrease in the level of RPE65 expression, a decrease in ocular retinoid levels concomitant with low visual function, and a rapid disorganization of RPE cells, ultimately leading to retinal degeneration. Our results identify the ERK1/2 pathway as a direct regulator of the visual cycle and a critical component of the viability of RPE and photoreceptor cells. Moreover, our results caution about the need for a very fine adjustment of kinase inhibition in cancer or ARMD treatment in order to avoid ocular side effects
Spatial evolution of magnetic reconnection diffusion region structures with distance from the X-line
No full textWe report Magnetospheric Multiscale four-spacecraft observations of a thin reconnecting current sheet with weakly asymmetric inflow conditions and a guide field of approximately twice the reconnecting magnetic field. The event was observed at the interface of interlinked magnetic field lines at the flank magnetopause when the maximum spacecraft separation was 370 km and the spacecraft covered & SIM;1.7 ion inertial lengths (d(i)) in the reconnection outflow direction. The ion-scale spacecraft separation made it possible to observe the transition from electron-only super ion-Alfvenic outflow near the electron diffusion region (EDR) to the emergence of sub-Alfvenic ion outflow in the ion diffusion region (IDR). The EDR to IDR evolution over a distance less than 2 d(i) also shows the transition from a near-linear reconnecting magnetic field reversal to a more bifurcated current sheet as well as significant decreases in the parallel electric field and dissipation. Both the ion and electron heating in this diffusion region event were similar to the previously reported heating in the far downstream exhausts. The dimensionless reconnection rate, obtained four different ways, was in the range of 0.13-0.27. This event reveals the rapid spatial evolution of the plasma and electromagnetic fields through the EDR to IDR transition region.& nbsp;(C) 2021 Author(s). All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/)
Trichoplein/mitostatin regulates endoplasmic reticulum-mitochondria juxtaposition
No full textTrichoplein/mitostatin (TpMs) is a keratin-binding protein that partly colocalizes with mitochondria and is often downregulated in epithelial cancers, but its function remains unclear. In this study, we report that TpMs regulates the tethering between mitochondria and endoplasmic reticulum (ER) in a Mitofusin 2 (Mfn2)-dependent manner. Subcellular fractionation and immunostaining show that TpMs is present at the interface between mitochondria and ER. The expression of TpMs leads to mitochondrial fragmentation and loosens tethering with ER, whereas its silencing has opposite effects. Functionally, the reduced tethering by TpMs inhibits apoptosis by Ca2 +-dependent stimuli that require ERmitochondria juxtaposition. Biochemical and genetic evidence support a model in which TpMs requires Mfn2 to modulate mitochondrial shape and tethering. Thus, TpMs is a new regulator of mitochondriaER juxtaposition
