Identification of Fis1 Interactors in Toxoplasma gondii Reveals a Novel Protein Required for Peripheral Distribution of the Mitochondrion

Toxoplasma gondii’s single mitochondrion is very dynamic and undergoes morphological changes throughout the parasite’s life cycle. During parasite division, the mitochondrion elongates, enters the daughter cells just prior to cytokinesis, and undergoes fission. Extensive morphological changes also occur as the parasite transitions from the intracellular environment to the extracellular environment. We show that treatment with the ionophore monensin causes reversible constriction of the mitochondrial outer membrane and that this effect depends on the function of the fission-related protein Fis1. We also observed that mislocalization of the endogenous Fis1 causes a dominant-negative effect that affects the morphology of the mitochondrion. As this suggests that Fis1 interacts with proteins critical for maintenance of mitochondrial structure, we performed various protein interaction trap screens. In this manner, we identified a novel outer mitochondrial membrane protein, LMF1, which is essential for positioning of the mitochondrion in intracellular parasites. Normally, while inside a host cell, the parasite mitochondrion is maintained in a lasso shape that stretches around the parasite periphery where it has regions of coupling with the parasite pellicle, suggesting the presence of membrane contact sites. In intracellular parasites lacking LMF1, the mitochondrion is retracted away from the pellicle and instead is collapsed, as normally seen only in extracellular parasites. We show that this phenotype is associated with defects in parasite fitness and mitochondrial segregation. Thus, LMF1 is necessary for mitochondrial association with the parasite pellicle during intracellular growth, and proper mitochondrial morphology is a prerequisite for mitochondrial division

Characterization of a Novel Fis1 Interactor Required for Peripheral Distribution of the Mitochondrion of Toxoplasma Gondii

Indiana University-Purdue University Indianapolis (IUPUI)Toxoplasma’s singular mitochondrion is extremely dynamic and undergoes morphological changes throughout the parasite’s life cycle. While intracellular‚ the mitochondrion is maintained in a lasso shape that stretches around the parasite periphery and is in close proximity to the pellicle‚ suggesting the presence of membrane contact sites. Upon egress‚ these contact sites disappear‚ and the mitochondrion retracts and collapses towards the apical end of the parasite. Once reinvaded‚ the lasso shape is quickly reformed‚ indicating that dynamic membrane contact sites regulate the positioning of the mitochondrion. We discovered a novel protein (TgGT1_265180) that associates with the mitochondrion via interactions with the fission related protein Fis1. Knockout of TgGT1_265180‚ which we have dubbed LMF1 for Lasso Maintenance Factor 1‚ results in a complete disruption of the normal mitochondrial morphology. In intracellular LMF1 knockout parasites, the mitochondrial lasso shape is disrupted‚ and instead it is collapsed as normally only seen in extracellular parasites. Additionally, proper mitochondrial segregation is disrupted‚ resulting in parasites with no mitochondrion and extra mitochondrial material outside of the parasites. These gross morphological changes are associated with a significant reduction of parasite propagation and can be rescued by reintroduction of a wildtype copy of LMF1. Co-immunoprecipitations and Yeast Two-Hybrid predict interactions with the parasite pellicle. Therefore, we hypothesize that LMF1 mediates contact between the mitochondrion and the pellicle in a regulatable fashion‚ and that the LMF1-dependent morphodynamics are critical for parasite propagation. Current studies are focused on characterizing the consequences of mitochondrial collapse and identifying proteins that interact with LMF1 to position the mitochondrion to the periphery of the parasite

TgDrpC, an atypical dynamin‐related protein in Toxoplasma gondii, is associated with vesicular transport factors and parasite division

Dynamin‐related proteins (Drps) are involved in diverse processes such as organelle division and vesicle trafficking. The intracellular parasite Toxoplasma gondii possesses three distinct Drps. TgDrpC, whose function remains unresolved, is unusual in that it lacks a conserved GTPase Effector Domain, which is typically required for function. Here, we show that TgDrpC localizes to cytoplasmic puncta; however, in dividing parasites, TgDrpC redistributes to the growing edge of the daughter cells. By conditional knockdown, we determined that loss of TgDrpC stalls division and leads to rapid deterioration of multiple organelles and the IMC. We also show that TgDrpC interacts with proteins that exhibit homology to those involved in vesicle transport, including members of the adaptor complex 2. Two of these proteins, a homolog of the adaptor protein 2 (AP‐2) complex subunit alpha‐1 and a homolog of the ezrin–radixin–moesin (ERM) family proteins, localize to puncta and associate with the daughter cells. Consistent with the association with vesicle transport proteins, re‐distribution of TgDrpC to the IMC during division is dependent on post‐Golgi trafficking. Together, these results support that TgDrpC contributes to vesicle trafficking and is critical for stability of parasite organelles and division

The Two Faces of Capacitance: New Interpretations for Electrical Impedance Measurements of Perovskite Solar Cells and Their Relation to Hysteresis

Perovskite solar cells are notorious for exhibiting transient behaviour not seen in conventional inorganic semiconductor devices. Significant inroads have been made into understanding this fact in terms of rapid ion migration, now a well-established property of the prototype photovoltaic perovskite MAPbI$_3$ and strongly implicated in the newer mixed compositions. Here we study the manifestations of ion migration in frequency-domain small-signal measurements, focusing on the popular technique of Electrical Impedance Spectroscopy (EIS). We provide new interpretations for a variety of previously puzzling features, including giant photo-induced low-frequency capacitance and negative capacitance in a variety of forms. We show that these apparently strange measurements can be rationalized by the splitting of AC current into two components, one associated with charge-storage, and the other with the quasi-steady-state recombination current of electrons and holes. The latter contribution to the capacitance can take either a positive or a negative sign, and is potentially very large when slow, voltage-sensitive processes such as ion migration are at play. Using numerical drift-diffusion semiconductor models, we show that giant photo-induced capacitance, inductive loop features, and low-frequency negative capacitance all emerge naturally as consequences of ion migration via its coupling to quasi-steady-state electron and hole currents. In doing so, we unify the understanding of EIS measurements with the comparably well-developed theory of rate dependent current-voltage (I-V) measurements in perovskite cells. Comparing the two techniques, we argue that EIS is more suitable for quantifying I-V hysteresis than conventional methods based on I-V sweeps, and demonstrate this application on a variety of cell types.Comment: Fixed typos and amended the axes on Figure 3 for clarit

Loss of Nmp4 optimizes osteogenic metabolism and secretion to enhance bone quality

A goal of osteoporosis therapy is to restore lost bone with structurally sound tissue. Mice lacking the transcription factor Nuclear Matrix Protein 4 (Nmp4, Zfp384, Ciz, ZNF384) respond to several classes of osteoporosis drugs with enhanced bone formation compared to wild type (WT) animals. Nmp4-/- mesenchymal stem/progenitor cells (MSPCs) exhibit an accelerated and enhanced mineralization during osteoblast differentiation. To address the mechanisms underlying this hyper-anabolic phenotype, we carried out RNA-sequencing and molecular and cellular analyses of WT and Nmp4-/- MSPCs during osteogenesis to define pathways and mechanisms associated with elevated matrix production. We determined that Nmp4 has a broad impact on the transcriptome during osteogenic differentiation, contributing to the expression of over 5,000 genes. Phenotypic anchoring of transcriptional data was performed for the hypothesis-testing arm through analysis of cell metabolism, protein synthesis and secretion, and bone material properties. Mechanistic studies confirmed that Nmp4-/- MSPCs exhibited an enhanced capacity for glycolytic conversion- a key step in bone anabolism. Nmp4-/- cells showed elevated collagen translation and secretion. Expression of matrix genes that contribute to bone material-level mechanical properties were elevated in Nmp4-/- cells, an observation that was supported by biomechanical testing of bone samples from Nmp4-/- and WT mice. We conclude that loss of Nmp4 increases the magnitude of glycolysis upon the metabolic switch, which fuels the conversion of the osteoblast into a super-secretor of matrix resulting in more bone with improvements in intrinsic quality

Complete genome sequence of phi29-like Microbacterium foliorum podovirus phage PineapplePizza

Bacteriophage PineapplePizza is a podovirus infecting Microbacterium foliorum NRRL B-24224. The genome is 16,662 bp long and contains 23 predicted protein-coding genes. Interestingly, PineapplePizza shows amino acid similarities to well-studied Bacillus subtilis phage phi29

In situ recombination junction between p-Si and TiO2 enables high-efficiency monolithic perovskite/Si tandem cells

Increasing the power conversion efficiency of silicon (Si) photovoltaics is a key enabler for continued reductions in the cost of solar electricity. Here, we describe a two-terminal perovskite/Si tandem design that increases the Si cell’s output in the simplest possible manner: by placing a perovskite cell directly on top of the Si bottom cell. The advantageous omission of a conventional interlayer eliminates both optical losses and processing steps and is enabled by the low contact resistivity attainable between n-type TiO2 and Si, established here using atomic layer deposition. We fabricated proof-of-concept perovskite/Si tandems on both homojunction and passivating contact heterojunction Si cells to demonstrate the broad applicability of the interlayer-free concept. Stabilized efficiencies of 22.9 and 24.1% were obtained for the homojunction and passivating contact heterojunction tandems, respectively, which could be readily improved by reducing optical losses elsewhere in the device. This work highlights the potential of emerging perovskite photovoltaics to enable low-cost, high-efficiency tandem devices through straightforward integration with commercially relevant Si solar cells

Monolithic Perovskite/Si Tandem Solar Cells: Pathways to Over 30% Efficiency

The article commences with a review focusing on three critical aspects of the perovskite/Si tandem technology: the evolution of efficiencies to date, comparisons of Si subcell choices, and the interconnection design strategies. Building on this review, a clear route is provided for minimizing optical losses aided by optical simulations of a recently reported high‐efficiency perovskite/Si tandem system, optimizations which result in tandem current densities of ≈20 mAcm−2 with front‐side texture. The primary focus is on electrical modeling on the Si‐subcell, in order to understand the efficiency potential of this cell under filtered light in a tandem configuration. The possibility of increasing the Si subcell efficiency by 1% absolute is offered through joint improvements to the bulk lifetime, which exceeds 4 ms, and improves surface passivation quality to saturation current densities below 10 fA cm−2. Polycrystalline‐Si/SiOx passivating contacts are proposed as a promising alternative to partial‐area rear contacts, with the potential for further simplifying cell fabrication and improving device performance. A combination of optical modeling of the complete tandem structure alongside electrical modeling of the Si‐subcell, both with state‐of‐the‐art modeling tools, provides the first complete picture of the practical efficiency potential of perovskite/Si tandems.This work was supported by the Australian Government through the Australian Renewable Energy Agency (ARENA), Grant MG005, and the Australian Research Council through Grant FT13010916

Interface passivation using ultrathin polymer–fullerene films for high-efficiency perovskite solar cells with negligible hysteresis

Interfacial carrier recombination is one of the dominant loss mechanisms in high efficiency perovskite solar cells, and has also been linked to hysteresis and slow transient responses in these cells. Here we demonstrate an ultrathin passivation layer consisting of a PMMA:PCBM mixture that can effectively passivate defects at or near to the perovskite/TiO2 interface, significantly suppressing interfacial recombination. The passivation layer increases the open circuit voltage of mixed-cation perovskite cells by as much as 80 mV, with champion cells achieving Voc ∼ 1.18 V. As a result, we obtain efficient and stable perovskite solar cells with a steady-state PCE of 20.4% and negligible hysteresis over a large range of scan rates. In addition, we show that the passivated cells exhibit very fast current and voltage response times of less than 3 s under cyclic illumination. This new passivation approach addresses one of the key limitations of current perovskite cells, and paves the way to further efficiency gains through interface engineering.Australian Renewable Energy Agency; Australian Research Council; MSTC (Grant No. 2016YFA0301300), NNSFC (Grant No. 11674402) and GSTP (Grant No. 201607010044, 201607020023

Efficient Passivation and Low Resistivity for p+-Si/TiO2Contact by Atomic Layer Deposition

The monolithic, two-terminal (2-T) perovskite/silicon tandem solar cell is a promising candidate to increase the power conversion efficiency beyond the theoretical limit of 29.4% for crystalline silicon solar cells. To achieve a high-efficiency 2-T tandem, it is critical to have an interface that can connect the bottom and top subcells together so that both efficient passivation and good electrical contact are achieved. The majority of works done to date in this area, applied an intermediate layer as the recombination layer between perovskite and silicon, which incurs higher manufacturing costs and an additional processing step. Here we demonstrate a unique and straightforward interlayer-free approach to passivating highly boron-doped low-resistivity n-Si using a thin layer of TiO2 fabricated by atomic layer deposition (ALD) and a suitable pretreatment of the silicon surface. The passivation of this film is found to be superior to that of thermally grown SiO2 formed at high temperatures over 700 °C. The TiOX layer leads to a sufficiently low contact resistance of 0.45 Ω.cm2 and high-quality passivation with a recombination current density (J0) of 152 fA/cm2. The structure is applicable to both perovskite/Si tandems and single-junction Si solar cells.We acknowledge the support of the Australian Renewable Energy Agency (ARENA), the Australian Centre for Advanced Photovoltaics (ACAP), and the ANFF ACT Node in carrying out this research. H.T.N. acknowledges the fellowship support of the Australian Centre for Advanced Photovoltaics. T.P.W. is the recipient of an Australian Research Council Future Fellowship (Project No. FT180100302) funded by the Australian Government