Revisiting regulatory roles of replication protein A in plant DNA metabolism.

MAIN CONCLUSION: This review provides insight into the roles of heterotrimeric RPA protein complexes encompassing all aspects of DNA metabolism in plants along with specific function attributed by individual subunits. It highlights research gaps that need further attention. Replication protein A (RPA), a heterotrimeric protein complex partakes in almost every aspect of DNA metabolism in eukaryotes with its principle role being a single-stranded DNA-binding protein, thereby providing stability to single-stranded (ss) DNA. Although most of our knowledge of RPA structure and its role in DNA metabolism is based on studies in yeast and animal system, in recent years, plants have also been reported to have diverse repertoire of RPA complexes (formed by combination of different RPA subunit homologs arose during course of evolution), expected to be involved in plethora of DNA metabolic activities. Here, we have reviewed all studies regarding role of RPA in DNA metabolism in plants. As combination of plant RPA complexes may vary largely depending on number of homologs of each subunit, next step for plant biologists is to develop specific functional methods for detailed analysis of biological roles of these complexes, which we have tried to formulate in our review. Besides, complete absence of any study regarding regulatory role of posttranslational modification of RPA complexes in DNA metabolism in plants, prompts us to postulate a hypothetical model of same in light of information from animal system. With our review, we envisage to stimulate the RPA research in plants to shift its course from descriptive to functional studies, thereby bringing a new angle of studying dynamic DNA metabolism in plants.

A facile route to transfer Cu nanoparticles to organic medium for better stabilization and improved photocatalytic activity towards N-formylation reaction.

Cu nanoparticles were prepared in an aqueous phase by means of a simple reduction-route using sodium borohydride as the reducing agent in the presence of ascorbic acid and polyvinylpyrrolidone (PVP). The hydrosol of the Cu nanoparticles deteriorated within a day. It compelled to initiate a scheme to stabilize the nanoparticles for a long period of time. Phase transfer to organic solvents using Benzyldimethylstearylammonium chloride (BDSAC) as a phase transfer agent was found to be an effective path in this respect. BDSAC performed the dual role of dragging the Cu nanoparticles from water to organic solvent and also acted as a capping agent along with PVP for better stabilization of Cu nanoparticles. The organosol of the Cu nanoparticles exhibited excellent stability and promising catalytic activity towards N-formylation reactions on a number of amine substrates in presence of visible green LED light. The yield and reusability of the catalyst were promising. All the samples were thoroughly characterized by UV-visible spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy, energy dispersive analysis of x-rays, x-ray photoelectron spectroscopy and thermo gravimetric analysis.

Tip variant of focal segmental glomerulosclerosis: is it truly a benign variant?

BACKGROUND: Even though frequently described as a benign entity, the outcomes of the tip variant of focal segmental glomerulosclerosis (FSGS) have proven to be unclear. METHODS: This retrospective study includes a cohort of tip variant cases who presented to us from 2009 to 2012 and the analysis of their presenting clinical, histopathological features and treatment outcomes in comparison to the not otherwise specified (NOS) variants from our center in East India. RESULTS: Of the 224 biopsies of primary FSGS, 30 cases were the tip variant (13.39%). The mean age of presentation was around 29 years, with 57% being males. A nephrotic presentation was seen in 87% of cases, with 20% showing a presentation at <18 years of age for the first time. Global sclerosis, interstitial fibrosis, tubular atrophy and arteriolar hyalinosis were seen more commonly in the NOS variant. Twenty five patients of tip variant received steroid therapy and eight received alternative immunosuppression. Around 87% of the tip variant cases achieved some form of remission in proteinuria and 13.3% had a doubling of creatinine at a median follow-up of 2 years in comparison to NOS group in which 80% achieved some form of remission and 20% had a doubling of creatinine. CONCLUSION: Though the histopathological features and treatment responsiveness of the tip variant appear to be better than the NOS variety, the prognostic outcome does not seem to be as favorable as implicated previously with an important percentage of patients showing progressive worsening of renal function within a relatively short time span (2 years) in our cohort.

Optimizing Surgical Management of Tibial Plateau Fractures: A Comparative Study of Minimally Invasive Versus Open Reduction Techniques.

BACKGROUND: Tibial plateau fractures pose a significant challenge to orthopedic surgeons due to their complex nature and potential for long-term morbidity. Surgical intervention is often necessary to restore anatomical alignment and optimize functional outcomes. This study aimed to evaluate the efficacy of minimally invasive percutaneous plate osteosynthesis (MIPPO) compared to open reduction and internal fixation (ORIF) in the management of tibial plateau fractures. MATERIALS AND METHODS: The present hospital-based observational study was conducted at Agartala Government Medical College for two years. Seventy adult patients with tibial plateau fractures were included, with surgical interventions performed based on fracture characteristics. Postoperative outcomes, including knee range of movements, functional recovery, and complication rates, were assessed at six months. RESULTS: MIPPO demonstrated superior outcomes compared to ORIF, with a higher proportion of participants achieving knee range of movements > 120 degrees (66.7% versus 36%; p = 0.030), excellent functional outcomes (66.7% versus 36%; p = 0.046), and lower postoperative complication rates (2.2% versus 28%; p = 0.001). Fracture union times were significantly shorter in MIPPO (12.49 weeks) when compared to ORIF (14 weeks) (p = 0.009). CONCLUSION: MIPPO offers advantages over conventional ORIF in terms of functional recovery and complication rates while demonstrating comparable fracture union times. These findings advocate for the adoption of MIPPO as a preferred surgical technique for tibial plateau fractures.

Visible-Light-Driven Mentha spicata L.-Mediated Ag-Doped Bi2Zr2O7 Nanocomposite for Enhanced Degradation of Organic Pollutants, Electrochemical Sensing, and Antibacterial Applications.

Novel visible-light-driven Ag (X)-doped Bi2Zr2O7 (BZO) nanocomposites in pudina (P) extract (Mentha spicata L.), X-1, 3, 5, 7, and 9 mol %, were synthesized by the one-pot greener solution combustion method. The as-synthesized nanocomposite materials were characterized by using various spectral [X-ray diffraction (XRD), Fourier transform infrared, UV-visible, UV- diffuse reflectance spectra, X-ray photoelectron spectroscopy], electrochemical (cyclic voltammetry, electrochemical impedance spectroscopy), and analytical (scanning electron microscopy-energy-dispersive X-ray spectroscopy, transmission electron microscopy, Brunauer-Emmett-Teller) techniques. The average particle size of the nanocomposite material was found to be between 14.8 and 39.2 nm by XRD. The well-characterized Ag-doped BZOP nanocomposite materials exhibited enhanced photocatalytic degradation activity toward hazardous dyes such as methylene blue (MB) and rose bengal (RB) under visible light irradiation ranges between 400 and 800 nm due to their low energy band gap. As a result, 7 mol % of Ag-doped BZOP nanocomposite material exhibited excellent photodegradation activity against MB (D.E. = 98.7%) and RB (D.E. = 99.3%) as compared to other Ag-doped BZOP nanocomposite materials and pure BZOP nanocomposite, respectively, due to enhanced semiconducting and optical behaviors, high binding energy, and mechanical and thermal stabilities. The Ag-doped BZOP nanocomposite material-based electrochemical sensor showed good sensing ability toward the determination of lead nitrate and dextrose with the lowest limit of detection (LOD) of 18 µM and 12 µM, respectively. Furthermore, as a result of the initial antibacterial screening study, the Ag-doped BZOP nanocomposite material was found to be more effective against Gram-negative bacteria (Escherichia coli) as compared to Gram-positive (Staphylococcus aureus) bacteria. The scavenger study reveals that radicals such as O2•- and •OH are responsible for MB and RB mineralization. TOC removal percentages were found to be 96.8 and 98.5% for MB and RB dyes, and experimental data reveal that the Ag-doped BZOP enhances the radical (O2•- and •OH) formation and MB and RB degradation under visible-light irradiation.

Defective mitochondrial COX1 translation due to loss of COX14 function triggers ROS-induced inflammation in mouse liver.

Mitochondrial oxidative phosphorylation (OXPHOS) fuels cellular ATP demands. OXPHOS defects lead to severe human disorders with unexplained tissue specific pathologies. Mitochondrial gene expression is essential for OXPHOS biogenesis since core subunits of the complexes are mitochondrial-encoded. COX14 is required for translation of COX1, the central mitochondrial-encoded subunit of complex IV. Here we describe a COX14 mutant mouse corresponding to a patient with complex IV deficiency. COX14M19I mice display broad tissue-specific pathologies. A hallmark phenotype is severe liver inflammation linked to release of mitochondrial RNA into the cytosol sensed by RIG-1 pathway. We find that mitochondrial RNA release is triggered by increased reactive oxygen species production in the deficiency of complex IV. Additionally, we describe a COA3Y72C mouse, affected in an assembly factor that cooperates with COX14 in early COX1 biogenesis, which displays a similar yet milder inflammatory phenotype. Our study provides insight into a link between defective mitochondrial gene expression and tissue-specific inflammation.

Defective COX1 expression in aging mice liver.

Mitochondrial defects are associated with aging processes and age-related diseases, including cardiovascular diseases, neurodegenerative diseases and cancer. In addition, some recent studies suggest mild mitochondrial dysfunctions appear to be associated with longer lifespans. In this context, liver tissue is considered to be largely resilient to aging and mitochondrial dysfunction. Yet, in recent years studies report dysregulation of mitochondrial function and nutrient sensing pathways in ageing livers. Therefore, we analyzed the effects of the aging process on mitochondrial gene expression in liver using wildtype C57BL/6N mice. In our analyses, we observed alteration in mitochondrial energy metabolism with age. To assess if defects in mitochondrial gene expression are linked to this decline, we applied a Nanopore sequencing based approach for mitochondrial transcriptomics. Our analyses show that a decrease of the Cox1 transcript correlates with reduced respiratory complex IV activity in older mice livers.

Synergistic effect of a Bi2Zr2O7 and hydroxyapatite composite: organic pollutant remediation, antibacterial and electrochemical sensing applications.

Global concern regarding the energy crisis and environmental pollution is increasing. The fabrication of efficient catalysts remains a long-term goal. Recently, green synthesis methods for catalyst fabrication have attracted the scientific community. Herein, a simple approach to synthesize bismuth zirconate-hydroxyapatite (BZO-HA) nanocomposites using Mentha spicata (mint) leaves as a reducing agent via a combustion method has been reported. The use of a green reducing agent provided economic attributes to this work. Among the prepared samples, the BZO-HA (20%) composite exhibited superior photocatalytic activity. The photodegradation efficiency of the composite reached 90.3% and 98.4% for methylene blue (MB) and rose Bengal (RB) dyes, respectively. The results showed the excellent optical performance of the prepared composites. The constructed sensor (BZO-HA 20%) for the very first time showed outstanding selectivity and performance towards sensing lead nitrate and dextrose compared to bare bismuth zirconate (BZO) and hydroxyapatite (HA). A three-electrode system using 0.1 M KCl was used for the study. The synthesized composite BZO-HA (20%) can sense lead nitrate and dextrose over the concentration range of 1-5 mM in the potential range from -1.0 V to +1.0 V. The BZO-HA composite was also investigated against Gram-negative (S. typhi) and Gram-positive (S. aureus) bacteria for antibacterial activity studies. Enhanced antibacterial activity was observed compared to bare BZO and HA catalysts. Thus, the prepared BZO-HA nanocomposite exhibited multifunctional applications.

Green-engineered synthesis of Bi2Zr2O7 NPs: excellent performance on electrochemical sensor and sunlight-driven photocatalytic studies.

In this rapid growing eco-friendly research world, synthesis of non-toxic, highly effective photocatalyst for potential applications is necessary. Herein, a strong ability Bi2Zr2O7 nanoparticle (BZO NP) with pyrochlore structure was fabricated by solution combustion synthesis using green (Mentha spicata) and chemical (Glycine) fuels. The X-ray diffraction analysis confirms the formation of pure phase for synthesized BZO NP using pudina extract (BZOP NP) compared to BZO NP using Glycine fuel (BZOG NP). The lower energy band gap of synthesized BZOP NP was observed than BZOG NP and its values were found to be 2.26 and 2.49 eV measured by UV-visible absorbance spectral technique. The morphological analysis of pores and voids formation as examined by scanning electron microscopy (SEM) technique. The synthesized BZOP NP shows excellent photocatalytic activity for degradation of three different dyes under sunlight irradiation for about 150 min with 97.9% for Rose Bengal (RB) dye with lower charge transfer resistance (Rct) value. For the very first time, the synthesized NPs can be utilized as supercapacitor with good specific capacitance (SPCcv) value of 14.3 F/g and SPCGD (12.5 F/g) for BZOP compared to BZOG indicating pseudocapacitance nature. The synthesized nanoparticles (NPs) can sense lead nitrate and dextrose at concentration 1-5 mM in the potential range of - 1.0 to + 1.0 V. Accordingly, the reduction potential peak at - 0.25 V and oxidation potential peak found at - 0.82 V confirms the presence of lead content and presence of additional potential peaks at - 0.37 V and - 0.71 V for detection of dextrose biochemical. Recyclability experiment showed the retainment of photocatalytic activity up to five cycles indicating the photostability.

Metabolic switch from fatty acid oxidation to glycolysis in knock-in mouse model of Barth syndrome.

Mitochondria are central for cellular metabolism and energy supply. Barth syndrome (BTHS) is a severe disorder, due to dysfunction of the mitochondrial cardiolipin acyl transferase tafazzin. Altered cardiolipin remodeling affects mitochondrial inner membrane organization and function of membrane proteins such as transporters and the oxidative phosphorylation (OXPHOS) system. Here, we describe a mouse model that carries a G197V exchange in tafazzin, corresponding to BTHS patients. TAZG197V mice recapitulate disease-specific pathology including cardiac dysfunction and reduced oxidative phosphorylation. We show that mutant mitochondria display defective fatty acid-driven oxidative phosphorylation due to reduced levels of carnitine palmitoyl transferases. A metabolic switch in ATP production from OXPHOS to glycolysis is apparent in mouse heart and patient iPSC cell-derived cardiomyocytes. An increase in glycolytic ATP production inactivates AMPK causing altered metabolic signaling in TAZG197V . Treatment of mutant cells with AMPK activator reestablishes fatty acid-driven OXPHOS and protects mice against cardiac dysfunction.

IDH3γ functions as a redox switch regulating mitochondrial energy metabolism and contractility in the heart.

Redox signaling and cardiac function are tightly linked. However, it is largely unknown which protein targets are affected by hydrogen peroxide (H2O2) in cardiomyocytes that underly impaired inotropic effects during oxidative stress. Here, we combine a chemogenetic mouse model (HyPer-DAO mice) and a redox-proteomics approach to identify redox sensitive proteins. Using the HyPer-DAO mice, we demonstrate that increased endogenous production of H2O2 in cardiomyocytes leads to a reversible impairment of cardiac contractility in vivo. Notably, we identify the γ-subunit of the TCA cycle enzyme isocitrate dehydrogenase (IDH)3 as a redox switch, linking its modification to altered mitochondrial metabolism. Using microsecond molecular dynamics simulations and experiments using cysteine-gene-edited cells reveal that IDH3γ Cys148 and 284 are critically involved in the H2O2-dependent regulation of IDH3 activity. Our findings provide an unexpected mechanism by which mitochondrial metabolism can be modulated through redox signaling processes.

Role of Mitochondrial Nucleic Acid Sensing Pathways in Health and Patho-Physiology.

Mitochondria, in symbiosis with the host cell, carry out a wide variety of functions from generating energy, regulating the metabolic processes, cell death to inflammation. The most prominent function of mitochondria relies on the oxidative phosphorylation (OXPHOS) system. OXPHOS heavily influences the mitochondrial-nuclear communication through a plethora of interconnected signaling pathways. Additionally, owing to the bacterial ancestry, mitochondria also harbor a large number of Damage Associated Molecular Patterns (DAMPs). These molecules relay the information about the state of the mitochondrial health and dysfunction to the innate immune system. Consequently, depending on the intracellular or extracellular nature of detection, different inflammatory pathways are elicited. One group of DAMPs, the mitochondrial nucleic acids, hijack the antiviral DNA or RNA sensing mechanisms such as the cGAS/STING and RIG-1/MAVS pathways. A pro-inflammatory response is invoked by these signals predominantly through type I interferon (T1-IFN) cytokines. This affects a wide range of organ systems which exhibit clinical presentations of auto-immune disorders. Interestingly, tumor cells too, have devised ingenious ways to use the mitochondrial DNA mediated cGAS-STING-IRF3 response to promote neoplastic transformations and develop tumor micro-environments. Thus, mitochondrial nucleic acid-sensing pathways are fundamental in understanding the source and nature of disease initiation and development. Apart from the pathological interest, recent studies also attempt to delineate the structural considerations for the release of nucleic acids across the mitochondrial membranes. Hence, this review presents a comprehensive overview of the different aspects of mitochondrial nucleic acid-sensing. It attempts to summarize the nature of the molecular patterns involved, their release and recognition in the cytoplasm and signaling. Finally, a major emphasis is given to elaborate the resulting patho-physiologies.

A tale of monoclonal immunoglobulin: Clinicopathological analysis of proliferative glomerulonephritis with monoclonal immunoglobulin deposit.

BACKGROUND: Proliferative glomerulonephritis with monoclonal immunoglobulin deposit (PGNMID) is an entity with a variable clinical and histological spectrum, which mimics immune-complex mediated glomerulonephritis on light microscopy. In this article, we aim to describe the clinical and pathological features of six cases of PGNMID that we encountered during our routine practice. MATERIALS AND METHODS: The study was of the prospective type carried out from February 2018 to August 2019. The renal biopsies that we received in our department, were processed for light microscopy, immunofluorescence microscopy, and electron microscopy. Light microscopic findings were carefully re-evaluated by two experienced renal pathologists. Key diagnostic features were 1) Monoclonal staining of glomeruli for one immunoglobulin (Ig) subclass and single light chain, 2) Membranoproliferative glomerulonephritis (MPGN) pattern (rarely membranous or crescentic), 3) Subendothelial and mesangial (rarely subepithelial) deposits. RESULTS: : We diagnosed five cases of IgG PGNMID and one case of IgA PGNMID with a mean age 53 ± 10.33 years. The most common histological pattern, seen in three cases was MPGN. IgG3 deposits were identified in five cases out of which k light chain restriction was present in four cases and λ light chain restriction was present in one case. IgA deposits were identified in one case that had λ light chain restriction. One patient suffered from multiple myeloma. CONCLUSIONS: The renal biopsy especially immunofluorescence analysis is the key modality for diagnosis of PGNMID where it shows staining of the glomerulus for a single heavy-chain subclass and a single light-chain isotype. Electron microscopic evaluation is necessary to differentiate PGNMID from other renal diseases with monoclonal immunoglobulin deposits.

A tale of tripartite: An experience from a tertiary care center of Eastern India.

BACKGROUND: Crescentic glomerulonephritis (Cr GN) is pattern of glomerular injury resulting from wide range of diseases sharing a common pathogenesis. OBJECTIVES: The objective of our study was to analyze the clinicopathological spectrum and outcome of Cr GN with special reference to its immunopathological subtypes using a panel of immunofluorescence stains. MATERIALS AND METHODS: Native renal biopsies with crescentic pattern of injury were included. Detailed Clinical and laboratory variables were analyzed along with the treatment protocol and renal outcome, wherever available. Renal biopsy slides were evaluated for various glomerular and extraglomerular features. Both qualitative and quantitative data were analyzed. RESULTS: A total of 57 cases of Cr GN were included; majority (47.36%) of cases were pauci-immune in nature. Among clinical features, ranges of proteinuria and creatinine level were significantly different between subgroups. The various light microscopic parameters, including proportion of cellular crescents and capillary wall necrosis were different. Presence of arteriolar changes also showed association with unfavorable outcome. Three unusual associations, including IgA nephropathy, membranous glomerulonephritis and Hepatitis B infection were detected. Adequate follow-up information was available in 35 of the patients. Of these, 14 were dialysis-dependent at the last follow-up. CONCLUSIONS: Type III Cr GN (pauci-immune Cr GN) was the commonest cause of Cr GN in our population. Adult patients required renal replacement therapy more frequently than pediatric cases those are chiefly infection associated. Critical appraisal of clinical, histopathological and immunofluorescence finding help to identify individual subtypes as treatment and outcome varies accordingly.

Genetic analysis of HIV-1 Circulating Recombinant Form 02_AG, B and C subtype-specific envelope sequences from Northern India and their predicted co-receptor usage.

HIV-1 epidemic in India is largely driven by subtype C but other subtypes or recombinants have also been reported from several states of India. This is mainly due to the co-circulation of other genetic subtypes that potentially can recombine to generate recombinant/mosaic genomes. In this study, we report detail genetic characterization of HIV-1 envelope sequences from North India (Delhi and neighboring regions). Six of 13 were related to subtype C, one B and the rest six showed relatedness with CRF02_AG strain. The subtype C possessed the highly conserved GPGQ motif but subtype B possessed the GPGR motif in the V3 loop as observed earlier. While most of the sequences suggested CCR5 co-receptor usage, one subtype C sample clearly indicated CXCR4 usage. A successful mother to child transmission was established in two pairs. Thus, co-circulation of multiple subtypes (B and C) and the recombinant CRF02_AG strains in North India suggests a rapidly evolving scenario of HIV-1 epidemic in this region with impact on vaccine formulation. Since this is the first report of CRF02_AG envelope from India, it will be important to monitor the spread of this strain and its impact on HIV-1 transmission in India.

How Our Perception and Confidence Are Altered Using Decision Cues.

Understanding how individuals utilize social information while making perceptual decisions and how it affects their decision confidence is crucial in a society. To date, very little has been known about perceptual decision-making in humans and the associated neural mediators under social influence. The present study provides empirical evidence of how individuals are manipulated by others' decisions while performing a face/car identification task. Subjects were significantly influenced by what they perceived as the decisions of other subjects, while the cues, in reality, were manipulated independently from the stimulus. Subjects, in general, tend to increase their decision confidence when their individual decision and the cues coincide, while their confidence decreases when cues conflict with their individual judgments, often leading to reversal of decision. Using a novel statistical model, it was possible to rank subjects based on their propensity to be influenced by cues. This was subsequently corroborated by an analysis of their neural data. Neural time series analysis revealed no significant difference in decision-making using social cues in the early stages, unlike neural expectation studies with predictive cues. Multivariate pattern analysis of neural data alludes to a potential role of the frontal cortex in the later stages of visual processing, which appeared to code the effect of cues on perceptual decision-making. Specifically, the medial frontal cortex seems to play a role in facilitating perceptual decision preceded by conflicting cues.

Overexpression of branched-chain amino acid aminotransferases rescues the growth defects of cells lacking the Barth syndrome-related gene TAZ1.

The yeast protein Taz1 is the orthologue of human Tafazzin, a phospholipid acyltransferase involved in cardiolipin (CL) remodeling via a monolyso CL (MLCL) intermediate. Mutations in Tafazzin lead to Barth syndrome (BTHS), a metabolic and neuromuscular disorder that primarily affects the heart, muscles, and immune system. Similar to observations in fibroblasts and platelets from patients with BTHS or from animal models, abolishing yeast Taz1 results in decreased total CL amounts, increased levels of MLCL, and mitochondrial dysfunction. However, the biochemical mechanisms underlying the mitochondrial dysfunction in BTHS remain unclear. To better understand the pathomechanism of BTHS, we searched for multi-copy suppressors of the taz1Δ growth defect in yeast cells. We identified the branched-chain amino acid transaminases (BCATs) Bat1 and Bat2 as such suppressors. Similarly, overexpression of the mitochondrial isoform BCAT2 in mammalian cells lacking TAZ improves their growth. Elevated levels of Bat1 or Bat2 did not restore the reduced membrane potential, altered stability of respiratory complexes, or the defective accumulation of MLCL species in yeast taz1Δ cells. Importantly, supplying yeast or mammalian cells lacking TAZ1 with certain amino acids restored their growth behavior. Hence, our findings suggest that the metabolism of amino acids has an important and disease-relevant role in cells lacking Taz1 function. KEY MESSAGES: Bat1 and Bat2 are multi-copy suppressors of retarded growth of taz1Δ yeast cells. Overexpression of Bat1/2 in taz1Δ cells does not rescue known mitochondrial defects. Supplementation of amino acids enhances growth of cells lacking Taz1 or Tafazzin. Altered metabolism of amino acids might be involved in the pathomechanism of BTSH.

COX16 promotes COX2 metallation and assembly during respiratory complex IV biogenesis.

Cytochrome c oxidase of the mitochondrial oxidative phosphorylation system reduces molecular oxygen with redox equivalent-derived electrons. The conserved mitochondrial-encoded COX1- and COX2-subunits are the heme- and copper-center containing core subunits that catalyze water formation. COX1 and COX2 initially follow independent biogenesis pathways creating assembly modules with subunit-specific, chaperone-like assembly factors that assist in redox centers formation. Here, we find that COX16, a protein required for cytochrome c oxidase assembly, interacts specifically with newly synthesized COX2 and its copper center-forming metallochaperones SCO1, SCO2, and COA6. The recruitment of SCO1 to the COX2-module is COX16- dependent and patient-mimicking mutations in SCO1 affect interaction with COX16. These findings implicate COX16 in CuA-site formation. Surprisingly, COX16 is also found in COX1-containing assembly intermediates and COX2 recruitment to COX1. We conclude that COX16 participates in merging the COX1 and COX2 assembly lines.

Defective Mitochondrial Cardiolipin Remodeling Dampens HIF-1α Expression in Hypoxia.

Mitochondria fulfill vital metabolic functions and act as crucial cellular signaling hubs, integrating their metabolic status into the cellular context. Here, we show that defective cardiolipin remodeling, upon loss of the cardiolipin acyl transferase tafazzin, decreases HIF-1α signaling in hypoxia. Tafazzin deficiency does not affect posttranslational HIF-1α regulation but rather HIF-1α gene expression, a dysfunction recapitulated in iPSC-derived cardiomyocytes from Barth syndrome patients with tafazzin deficiency. RNA-seq analyses confirmed drastically altered signaling in tafazzin mutant cells. In hypoxia, tafazzin-deficient cells display reduced production of reactive oxygen species (ROS) perturbing NF-κB activation and concomitantly HIF-1α gene expression. Tafazzin-deficient mice hearts display reduced HIF-1α levels and undergo maladaptive hypertrophy with heart failure in response to pressure overload challenge. We conclude that defective mitochondrial cardiolipin remodeling dampens HIF-1α signaling due to a lack of NF-κB activation through reduced mitochondrial ROS production, decreasing HIF-1α transcription.

Fibulin-6 regulates pro-fibrotic TGF-ß responses in neonatal mouse ventricular cardiac fibroblasts.

Fibulin-6, an essential component of extracellular matrix determines the architecture of cellular junctions in tissues undergoing strain. Increased expression and deposition of fibulin-6 facilitates fibroblast migration in response to TGF-ß, following myocardial infarction in mouse heart. The underlying mechanism still remains elusive. In conjunction with our previous study, we have now demonstrated that in fibulin-6 knockdown (KD) fibroblasts, not only TGF-ß dependent migration, but also stress fiber formation, cellular networking and subsequently fibroblast wound contraction is almost abrogated. SMAD dependent TGF-ß pathway shows ~75% decreased translocation of R-SMAD and co-SMAD into the nucleus upon fibulin-6 KD. Consequently, SMAD dependent pro-fibrotic gene expression is considerably down regulated to basal levels both in mRNA and protein. Also, investigating the non-SMAD pathways we observed a constitutive increase in pERK-levels in fibulin-6 KD fibroblast compared to control, but no change was seen in pAKT. Immunoprecipitation studies revealed 60% reduced interaction of TGF-ß receptor II and I (TGFRII and I) accompanied by diminished phosphorylation of TGFRI at serin165 in fibulin-6 KD cells. In conclusion, fibulin-6 plays an important role in regulating TGF-ß mediated responses, by modulating TGF-ß receptor dimerization and activation to further trigger downstream pathways.