NDUFAF5 Hydroxylates NDUFS7 at an Early Stage in the Assembly of Human Complex I.

Complex I (NADH ubiquinone oxidoreductase) in mammalian mitochondria is an L-shaped assembly of 45 proteins. One arm lies in the inner membrane, and the other extends about 100 Å into the matrix of the organelle. The extrinsic arm contains binding sites for NADH, the primary electron acceptor FMN, and seven iron-sulfur clusters that form a pathway for electrons linking FMN to the terminal electron acceptor, ubiquinone, which is bound in a tunnel in the region of the junction between the arms. The membrane arm contains four antiporter-like domains, energetically coupled to the quinone site and involved in pumping protons from the matrix into the intermembrane space contributing to the proton motive force. Seven of the subunits, forming the core of the membrane arm, are translated from mitochondrial genes, and the remaining subunits, the products of nuclear genes, are imported from the cytosol. Their assembly is coordinated by at least thirteen extrinsic assembly factor proteins that are not part of the fully assembled complex. They assist in insertion of co-factors and in building up the complex from smaller sub-assemblies. One such factor, NDUFAF5, belongs to the family of seven-β-strand S-adenosylmethionine-dependent methyltransferases. However, similar to another family member, RdmB, it catalyzes the introduction of a hydroxyl group, in the case of NDUFAF5, into Arg-73 in the NDUFS7 subunit of human complex I. This modification occurs early in the pathway of assembly of complex I, before the formation of the juncture between peripheral and membrane arms.This work was supported by the Medical Research Council via Intramural Program U105663150 and Program Grant MR/M009858/1 (to J. E. W.)

Persistence of the mitochondrial permeability transition in the absence of subunit c of human ATP synthase.

The permeability transition in human mitochondria refers to the opening of a nonspecific channel, known as the permeability transition pore (PTP), in the inner membrane. Opening can be triggered by calcium ions, leading to swelling of the organelle, disruption of the inner membrane, and ATP synthesis, followed by cell death. Recent proposals suggest that the pore is associated with the ATP synthase complex and specifically with the ring of c-subunits that constitute the membrane domain of the enzyme's rotor. The c-subunit is produced from three nuclear genes, ATP5G1, ATP5G2, and ATP5G3, encoding identical copies of the mature protein with different mitochondrial-targeting sequences that are removed during their import into the organelle. To investigate the involvement of the c-subunit in the PTP, we generated a clonal cell, HAP1-A12, from near-haploid human cells, in which ATP5G1, ATP5G2, and ATP5G3 were disrupted. The HAP1-A12 cells are incapable of producing the c-subunit, but they preserve the characteristic properties of the PTP. Therefore, the c-subunit does not provide the PTP. The mitochondria in HAP1-A12 cells assemble a vestigial ATP synthase, with intact F1-catalytic and peripheral stalk domains and the supernumerary subunits e, f, and g, but lacking membrane subunits ATP6 and ATP8. The same vestigial complex plus associated c-subunits was characterized from human 143B ρ(0) cells, which cannot make the subunits ATP6 and ATP8, but retain the PTP. Therefore, none of the membrane subunits of the ATP synthase that are involved directly in transmembrane proton translocation is involved in forming the PTP.This work was supported by the Medical Research Council (MRC) of the United Kingdom by Grant MC_U1065663150 and by Programme Grant MR/M009858/1 (to J.E.W.). H.C.F. received an MRC PhD studentship

Human METTL20 methylates lysine residues adjacent to the recognition loop of the electron transfer flavoprotein in mitochondria.

In mammalian mitochondria, protein methylation is a relatively uncommon post-transcriptional modification, and the extent of the mitochondrial protein methylome, the modifying methyltransferases, and their substrates have been little studied. As shown here, the β-subunit of the electron transfer flavoprotein (ETF) is one such methylated protein. The ETF is a heterodimer of α- and β-subunits. Lysine residues 199 and 202 of mature ETFβ are almost completely trimethylated in bovine heart mitochondria, whereas ETFα is not methylated. The enzyme responsible for the modifications was identified as methyltransferase-like protein 20 (METTL20). In human 143B cells, the methylation of ETFβ is less extensive and is diminished further by suppression of METTL20. Tagged METTL20 expressed in HEK293T cells specifically associates with the ETF and promotes the trimethylation of ETFβ lysine residues 199 and 202. ETF serves as a mobile electron carrier linking dehydrogenases involved in fatty acid oxidation and one-carbon metabolism to the membrane-associated ubiquinone pool. The methylated residues in ETFβ are immediately adjacent to a protein loop that recognizes and binds to the dehydrogenases. Suppression of trimethylation of ETFβ in mouse C2C12 cells oxidizing palmitate as an energy source reduced the consumption of oxygen by the cells. These experiments suggest that the oxidation of fatty acids in mitochondria and the passage of electrons via the ETF may be controlled by modulating the protein-protein interactions between the reduced dehydrogenases and the β-subunit of the ETF by trimethylation of lysine residues. METTL20 is the first lysine methyltransferase to be found to be associated with mitochondria.This work was supported by the Medical Research Council (MRC), UK

Permeability transition in human mitochondria persists in the absence of peripheral stalk subunits of ATP synthase

The opening of a non-specific channel, known as the permeability transition pore (PTP), in the inner membranes of mitochondria, can be triggered by calcium ions, leading to swelling of the organelle, disruption of the inner membrane and ATP synthesis, and cell death. Pore opening can be inhibited by cyclosporin A mediated via cyclophilin D. It has been proposed that the pore is associated with the dimeric ATP synthase, and that the OSCP (oligomycin sensitivity conferral protein), a component of the enzyme’s peripheral stalk, provides the site where cyclophilin D interacts. Subunit b contributes a central α-helical structure to the peripheral stalk, extending from near the top of the enzyme’s catalytic domain and crossing the membrane domain of the enzyme via two α-helices. We investigated the possible involvement of the subunit b and the OSCP in the PTP by generating clonal cells, HAP1-Δb and HAP1-ΔOSCP, lacking the membrane domain of subunit b or the OSCP, respectively, in which the correponding genes ATP5F1 and ATP5O had been disrupted. Both cell lines preserve the characteristic properties of the PTP. Therefore, the membrane domain of subunit b does not contribute to the PTP, and the OSCP does not provide the site of interaction with cyclophilin D. The membrane subunits ATP6, ATP8 and subunit c have been eliminated previously from possible participation in the PTP. Therefore, the only subunits of ATP synthase that could participate in pore formation are e, f, g, DAPIT (diabetes associated protein in insulin sensitive tissues) and the 6.8 kDa proteolipid.This work was supported by Medical Research Council, United Kingdom Programme Grant MR/M009858/1 (to J.E.W.)

Human METTL12 is a mitochondrial methyltransferase that modifies citrate synthase

The protein methylome in mammalian mitochondria has been little studied until recently. Here, we describe that lysine-368 of human citrate synthase is methylated and that the modifying enzyme, localized in the mitochondrial matrix, is methyltransferase-like protein 12 (METTL12), a member of the family of 7β-strand methyltransferases. Lysine-368 is near the active site of citrate synthase, but removal of methylation has no effect on its activity. In mitochondria, it is possible that some or all of the enzymes of the citric acid cycle, including citrate synthase, are organized in metabolons to facilitate the channelling of substrates between participating enzymes. Thus, possible roles for the methylation of Lys-368 are in controlling substrate channelling itself, or in influencing protein–protein interactions in the metabolon.This work was supported by the Medical Research Council of the United Kingdom by grant MC_U1065663150 and by Programme Grant MR/M009858/1, both to JEW and a Fellowship from the Swiss Novartis Foundation to VFR

The Cosmic Lens All-Sky Survey: statistical strong lensing, cosmological parameters, and global properties of galaxy populations

Extensive analyses of statistical strong gravitational lensing are performed based on the final Cosmic Lens All Sky Survey (CLASS) well-defined statistical sample of flat spectrum radio sources and current estimates of galaxy luminosity functions per morphological type. The analyses are done under the assumption that galactic lenses are well-approximated by singular isothermal ellipsoids and early-type galaxies evolved passively since redshift $z \sim 1$. Depending on how the late-type galaxy population is treated (i.e., whether its characteristic velocity dispersion is constrained or not), we find for a flat universe with a cosmological constant that the present matter fraction of the present critical density $\Omega_{\rm m} = 0.31^{+0.27}_{-0.14}$ (68%) for the unconstrained case or $0.40^{+0.28}_{-0.16}$ (68%) for the constrained case, with an additional systematic uncertainty of $\approx 0.11$ arising from the present uncertainty in the distribution of CLASS sources in redshift and flux density. For a flat universe with a constant equation of state for dark energy w = $p_x$(pressure)/$\rho_x$(energy density), we find that $w < -0.55^{+0.18}_{-0.11}$ (68%) for the unconstrained case or $w < -0.41^{+0.28}_{-0.16}$ (68%) for the constrained case. For the equal frequencies of oblates and prolates, we find that $\sigma_{*}^{(e)} = 198^{+22}_{-18}$ km s$^{-1}$ (68%) for a `steep' $\alpha^{(e)}=-1$ or $\sigma_{*}^{(e)} = 181^{+18}_{-15}$ km s$^{-1}$ (68%) for a `shallow' $\alpha^{(e)}=-0.54$. Finally, from the relative frequencies of doubly-imaged sources and quadruply-imaged sources, we find that a mean projected mass ellipticity of early-type galaxies $\bar{\epsilon}_{\rm mass} = 0.42$ with a 68% lower limit of 0.28. (Abridged)Comment: 31 pages, 12figures, 6 tables, to appear in MNRAS (referee comments incorporated, a section on future prospects added

On the General Kerr/CFT Correspondence in Arbitrary Dimensions

We study conformal symmetries on the horizon of a general stationary and axisymmetric black hole. We find that there exist physically reasonable boundary conditions that uniquely determine a set of symmetry generators, which form one copy of the Virasoro algebra. For extremal black holes, Cardy's formula reproduces exactly the Bekenstein-Hawking entropy.Comment: 17 page

A Gene-Based Association Method for Mapping Traits Using Reference Transcriptome Data

Genome-wide association studies (GWAS) have identified thousands of variants robustly associated with complex traits. However, the biological mechanisms underlying these associations are, in general, not well understood. We propose a gene-based association method called PrediXcan that directly tests the molecular mechanisms through which genetic variation affects phenotype. The approach estimates the component of gene expression determined by an individual’s genetic profile and correlates ‘imputed’ gene expression with the phenotype under investigation to identify genes involved in the etiology of the phenotype. Genetically regulated gene expression is estimated using whole-genome tissue-dependent prediction models trained with reference transcriptome data sets. PrediXcan enjoys the benefits of gene-based approaches such as reduced multiple-testing burden and a principled approach to the design of follow-up experiments. Our results demonstrate that PrediXcan can detect known and new genes associated with disease traits and provide insights into the mechanism of these associations

f(R)f(R) gravity theories in the Palatini Formalism constrained from strong lensing

$f(R)$ gravity, capable of driving the late-time acceleration of the universe, is emerging as a promising alternative to dark energy. Various $f(R)$ gravity models have been intensively tested against probes of the expansion history, including type Ia supernovae (SNIa), the cosmic microwave background (CMB) and baryon acoustic oscillations (BAO). In this paper we propose to use the statistical lens sample from Sloan Digital Sky Survey Quasar Lens Search Data Release 3 (SQLS DR3) to constrain $f(R)$ gravity models. This sample can probe the expansion history up to $z\sim2.2$, higher than what probed by current SNIa and BAO data. We adopt a typical parameterization of the form $f(R)=R-\alpha H^2_0(-\frac{R}{H^2_0})^\beta$ with $\alpha$ and $\beta$ constants. For $\beta=0$ ($\Lambda$CDM), we obtain the best-fit value of the parameter $\alpha=-4.193$, for which the 95% confidence interval that is [-4.633, -3.754]. This best-fit value of $\alpha$ corresponds to the matter density parameter $\Omega_{m0}=0.301$, consistent with constraints from other probes. Allowing $\beta$ to be free, the best-fit parameters are $(\alpha, \beta)=(-3.777, 0.06195)$. Consequently, we give $\Omega_{m0}=0.285$ and the deceleration parameter $q_0=-0.544$. At the 95% confidence level, $\alpha$ and $\beta$ are constrained to [-4.67, -2.89] and [-0.078, 0.202] respectively. Clearly, given the currently limited sample size, we can only constrain $\beta$ within the accuracy of $\Delta\beta\sim 0.1$ and thus can not distinguish between $\Lambda$CDM and $f(R)$ gravity with high significance, and actually, the former lies in the 68% confidence contour. We expect that the extension of the SQLS DR3 lens sample to the SDSS DR5 and SDSS-II will make constraints on the model more stringent.Comment: 10 pages, 7 figures. Accepted for publication in MNRA

Uncovering treatment burden as a key concept for stroke care: a systematic review of qualitative research

&lt;b&gt;Background&lt;/b&gt; Patients with chronic disease may experience complicated management plans requiring significant personal investment. This has been termed ‘treatment burden’ and has been associated with unfavourable outcomes. The aim of this systematic review is to examine the qualitative literature on treatment burden in stroke from the patient perspective.&lt;p&gt;&lt;/p&gt; &lt;b&gt;Methods and findings&lt;/b&gt; The search strategy centred on: stroke, treatment burden, patient experience, and qualitative methods. We searched: Scopus, CINAHL, Embase, Medline, and PsycINFO. We tracked references, footnotes, and citations. Restrictions included: English language, date of publication January 2000 until February 2013. Two reviewers independently carried out the following: paper screening, data extraction, and data analysis. Data were analysed using framework synthesis, as informed by Normalization Process Theory. Sixty-nine papers were included. Treatment burden includes: (1) making sense of stroke management and planning care, (2) interacting with others, (3) enacting management strategies, and (4) reflecting on management. Health care is fragmented, with poor communication between patient and health care providers. Patients report inadequate information provision. Inpatient care is unsatisfactory, with a perceived lack of empathy from professionals and a shortage of stimulating activities on the ward. Discharge services are poorly coordinated, and accessing health and social care in the community is difficult. The study has potential limitations because it was restricted to studies published in English only and data from low-income countries were scarce.&lt;p&gt;&lt;/p&gt; &lt;b&gt;Conclusions&lt;/b&gt; Stroke management is extremely demanding for patients, and treatment burden is influenced by micro and macro organisation of health services. Knowledge deficits mean patients are ill equipped to organise their care and develop coping strategies, making adherence less likely. There is a need to transform the approach to care provision so that services are configured to prioritise patient needs rather than those of health care systems