Mitochondria and Aging: Dilution Is the Solution

Metabolic component depletion in model systems results in life-span extension, which has been difficult to reconcile with human metabolic pathologies. Recently, Rea et al. (2007) have shown that mitochondrial electron transport chain RNAi phenotypes in the worm C. elegans are dose dependent, providing an alternative view of mitochondrial function in longevity and metabolic diseases

The UPRmt Preserves Mitochondrial Import to Extend Lifespan

The mitochondrial unfolded protein response (UPRmt) is dedicated to promoting mitochondrial proteostasis and is linked to extreme longevity. The key regulator of this process is the transcription factor ATFS-1, which, upon UPRmt activation, is excluded from the mitochondria and enters the nucleus to regulate UPRmt genes. However, the repair proteins synthesized as a direct result of UPRmt activation must be transported into damaged mitochondria that had previously excluded ATFS-1 owing to reduced import efficiency. To address this conundrum, we analyzed the role of the import machinery when the UPRmt was induced. Using in vitro and in vivo analysis of mitochondrial proteins, we surprisingly find that mitochondrial import increases when the UPRmt is activated in an ATFS-1-dependent manner, despite reduced mitochondrial membrane potential. The import machinery is upregulated, and an intact import machinery is essential for UPRmt-mediated lifespan extension. ATFS-1 has a weak mitochondrial targeting sequence (MTS), allowing for dynamic subcellular localization during the initial stages of UPRmt activation

Live-Cell Microscopy Reveals Small Molecule Inhibitor Effects on MAPK Pathway Dynamics

Oncogenic mutations in the mitogen activated protein kinase (MAPK) pathway are prevalent in human tumors, making this pathway a target of drug development efforts. Recently, ATP-competitive Raf inhibitors were shown to cause MAPK pathway activation via Raf kinase priming in wild-type BRaf cells and tumors, highlighting the need for a thorough understanding of signaling in the context of small molecule kinase inhibitors. Here, we present critical improvements in cell-line engineering and image analysis coupled with automated image acquisition that allow for the simultaneous identification of cellular localization of multiple MAPK pathway components (KRas, CRaf, Mek1 and Erk2). We use these assays in a systematic study of the effect of small molecule inhibitors across the MAPK cascade either as single agents or in combination. Both Raf inhibitor priming as well as the release from negative feedback induced by Mek and Erk inhibitors cause translocation of CRaf to the plasma membrane via mechanisms that are additive in pathway activation. Analysis of Erk activation and sub-cellular localization upon inhibitor treatments reveals differential inhibition and activation with the Raf inhibitors AZD628 and GDC0879 respectively. Since both single agent and combination studies of Raf and Mek inhibitors are currently in the clinic, our assays provide valuable insight into their effects on MAPK signaling in live cells

Search for supersymmetry in final states with two same-sign or three leptons and jets using 36 fb⁻¹ of √s=13 TeV pp collision data with the ATLAS detector

A search for strongly produced supersymmetric particles using signatures involving multiple energetic jets and either two isolated same-sign leptons (e or μ), or at least three isolated leptons, is presented. The analysis relies on the identification of b-jets and high missing transverse momentum to achieve good sensitivity. A data sample of proton-proton collisions at s=13 TeV recorded with the ATLAS detector at the Large Hadron Collider in 2015 and 2016, corresponding to a total integrated luminosity of 36.1 fb −1 , is used for the search. No significant excess over the Standard Model prediction is observed. The results are interpreted in several simplified supersymmetric models featuring R-parity conservation or R-parity violation, extending the exclusion limits from previous searches. In models considering gluino pair production, gluino masses are excluded up to 1.87 TeV at 95% confidence level. When bottom squarks are pair-produced and decay to a chargino and a top quark, models with bottom squark masses below 700 GeV and light neutralinos are excluded at 95% confidence level. In addition, model-independent limits are set on a possible contribution of new phenomena to the signal region yields.[Figure not available: see fulltext.]

Observation of four-top-quark production in the multilepton final state with the ATLAS detector

This paper presents the observation of four-top-quark (tt¯tt¯) production in proton-proton collisions at the LHC. The analysis is performed using an integrated luminosity of 140 fb−1 at a centre-of-mass energy of 13 TeV collected using the ATLAS detector. Events containing two leptons with the same electric charge or at least three leptons (electrons or muons) are selected. Event kinematics are used to separate signal from background through a multivariate discriminant, and dedicated control regions are used to constrain the dominant backgrounds. The observed (expected) significance of the measured tt¯tt¯ signal with respect to the standard model (SM) background-only hypothesis is 6.1 (4.3) standard deviations. The tt¯tt¯ production cross section is measured to be 22.5+6.6−5.5 fb, consistent with the SM prediction of 12.0±2.4 fb within 1.8 standard deviations. Data are also used to set limits on the three-top-quark production cross section, being an irreducible background not measured previously, and to constrain the top-Higgs Yukawa coupling and effective field theory operator coefficients that affect tt¯tt¯ production

Observation of four-top-quark production in the multilepton final state with the ATLAS detector

This paper presents the observation of four-top-quark (tt¯tt¯ ) production in proton-proton collisions at the LHC. The analysis is performed using an integrated luminosity of 140 fb−1 at a centre-of-mass energy of 13 TeV collected using the ATLAS detector. Events containing two leptons with the same electric charge or at least three leptons (electrons or muons) are selected. Event kinematics are used to separate signal from background through a multivariate discriminant, and dedicated control regions are used to constrain the dominant backgrounds. The observed (expected) significance of the measured tt¯tt¯ signal with respect to the standard model (SM) background-only hypothesis is 6.1 (4.3) standard deviations. The tt¯tt¯ production cross section is measured to be 22.5+6.6−5.5 fb, consistent with the SM prediction of 12.0±2.4 fb within 1.8 standard deviations. Data are also used to set limits on the three-top-quark production cross section, being an irreducible background not measured previously, and to constrain the top-Higgs Yukawa coupling and effective field theory operator coefficients that affect tt¯tt¯ production

Erratum to: Observation of four-top-quark production in the multilepton final state with the ATLAS detector

No abstract available

Tissue-Specific Aspects of Mitochondria and Longevity in C. elegans

The connection between aging and metabolism seems obvious, but the particulars of this relationship remain obscure. Theories linking the two abound, centered on the fact that mitochondria are the location of much of the cell's free radical production and on the general correlation between lifespan and metabolic rate. The connection between longevity and mitochondrial function was strengthened by RNAi-based screens in the worm C. elegans, where RNAi knock-down of mitochondrial Electron Transport Chain (ETC) subunits of Complex I (nuo-2), Complex III (cyc-1), Complex IV (cco-1) and Complex V (atp-3) extended lifespan (Dillin et al., 2002b; Lee et al., 2002). The effects of ETC knockdown on life span appear to depend on a mechanism more complex than direct effects on mitochondrial metabolic rates. Instead, they point towards the existence of signaling programs emanating from the mitochondria and capable of regulating the life span of the entire organism. The identity of components and timing requirements of the mitochondrial ETC that influence longevity have led us to search for a mechanism imposed during early development that sets the rate of aging for the remainder of the animal's life cycle as well as the critical tissues required for the mitochondrial ETC to set the rate of aging. I have found that disruption of cco-1, in the neurons or intestine of the worm is sufficient to confer a longevity phenotype. This tissue-specific knockdown was able to uncouple some of the detrimental phenotypes of organism-wide knockdown, such as reduced brood size and slow movement. Furthermore, the long lifespan that results from ETC perturbance is dependent on the mitochondrial Unfolded Protein Response (UPRmt) gene, ubl-5. Knowing that there are tissues in which ETC function is critical as well as the necessity for a functional UPRmt might allow us to better understand the basis of mitochondria-mediated longevity

Neuroendocrine Coordination of Mitochondrial Stress Signaling and Proteostasis

During neurodegenerative disease, the toxic accumulation of aggregates and misfolded proteins is often accompanied with widespread changes in peripheral metabolism, even in cells in which the aggregating protein is not present. The mechanism by which the central nervous system elicits a distal reaction to proteotoxic stress remains unknown. We hypothesized that the endocrine communication of neuronal stress plays a causative role in the changes in mitochondrial homeostasis associated with proteotoxic disease states. We find that an aggregation-prone protein expressed in the neurons of C. elegans binds to mitochondria, eliciting a global induction of a mitochondrial-specific unfolded protein response (UPR(mt)), affecting whole-animal physiology. Importantly, dense core vesicle release and secretion of the neurotransmitter serotonin is required for the signal's propagation. Collectively, these data suggest the commandeering of a nutrient sensing network to allow for cell-to-cell communication between mitochondria in response to protein folding stress in the nervous system