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1.
J Phys Chem Lett ; 15(28): 7272-7279, 2024 Jul 18.
Artículo en Inglés | MEDLINE | ID: mdl-38979897

RESUMEN

The solar-blind ultraviolet band presents a unique opportunity for low-background-noise detection due to limited atmospheric light transmission. Especially, with the ability to obtain size, shape and position information on the objects, the solar-blind image sensors are receiving increasing attention. However, because the inhibition of the crosstalk in crossbar arrays induces the complexity of the preparation process, rarely are applicable solar-blind UV imaging arrays reported. This work prepared an amorphous gallium oxide (a-Ga2O3)-based diode with a remarkable rectifier ratio of up to 106. Then an imaging array was prepared by one-step method in that the diodes working as the detection elements and the switching elements inhibiting crosstalk were simultaneously deposited. Moreover, a 2 × 2 crossbar array clearly demonstrates the inhibition of the crosstalk. This work presents a simple and cost-effective method for preparing applicable solar-blind imaging arrays that inhibits crosstalk, promoting the practical application of the Ga2O3-based solar-blind UV detectors.

2.
ACS Appl Mater Interfaces ; 16(8): 10398-10406, 2024 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-38380978

RESUMEN

The rapid evolution of the Internet of Things has engendered increased requirements for low-cost, self-powered UV photodetectors. Herein, high-performance self-driven UV photodetectors are fabricated by designing asymmetric metal-semiconductor-metal structures on the high-quality large-area CsCu2I3 microwire arrays. The asymmetrical depletion region doubles the photocurrent and response speed compared to the symmetric structure device, leading to a high responsivity of 233 mA/W to 355 nm radiation. Notably, at 0 V bias, the asymmetric device produces an open-circuit voltage of 356 mV and drives to a short-circuit current of 372 pA; meanwhile, the switch ratio (Iph/Idark) reaches up to 103, indicating its excellent potential for detecting weak light. Furthermore, the device maintains stable responses throughout 10000 UV-light switch cycles, with negligible degradation even after 90-day storage in air. Our work establishes that CsCu2I3 is a good candidate for self-powered UV detection and thoroughly demonstrates its potential as a passive device.

3.
J Phys Chem Lett ; 14(47): 10509-10516, 2023 Nov 30.
Artículo en Inglés | MEDLINE | ID: mdl-37970815

RESUMEN

Polarization-sensitive detectors have significant applications in modern communication and information processing. In this study. We present a polarization-sensitive detector based on a MoTe2/WTe2 heterojunction, where WTe2 forms a favorable bandgap structure with MoTe2 after forming the heterojunction. This enhances the carrier separation efficiency and photoelectric response. We successfully achieved wide spectral detection ranging from visible to near-infrared light. Specifically, under zero bias, our photodetector exhibits a responsivity (R) of 0.6 A/W and a detectivity (D*) of 3.6 × 1013 Jones for 635 nm laser illumination. Moreover, the photoswitching ratio can approach approximately 6.3 × 105. Importantly, the polarization sensitivity can reach 3.5 (5.2) at 635 (1310) nm polarized light at zero bias. This study both unveils potential for utilizing MoTe2/WTe2 heterojunctions as polarization-sensitive detectors and provides novel insights for developing high-performance optoelectronic devices.

4.
Nat Commun ; 14(1): 5228, 2023 08 26.
Artículo en Inglés | MEDLINE | ID: mdl-37633971

RESUMEN

Small interference RNAs are the key components of RNA interference, a conserved RNA silencing or viral defense mechanism in many eukaryotes. In Drosophila melanogaster, Dicer-2 (DmDcr-2)-mediated RNAi pathway plays important roles in defending against viral infections and protecting genome integrity. During the maturation of siRNAs, two cofactors can regulate DmDcr-2's functions: Loqs-PD that is required for dsRNA processing, and R2D2 that is essential for the subsequent loading of siRNAs into effector Ago2 to form RISC complexes. However, due to the lack of structural information, it is still unclear whether R2D2 and Loqs-PD affect the functions of DmDcr-2 simultaneously. Here we present several cryo-EM structures of DmDcr-2/R2D2/Loqs-PD complex bound to dsRNAs with various lengths by the Helicase domain. These structures revealed that R2D2 and Loqs-PD can bind to different regions of DmDcr-2 without interfering with each other. Furthermore, the cryo-EM results demonstrate that these complexes can form large oligomers and assemble into fibers. The formation and depolymerization of these oligomers are associated with ATP hydrolysis. These findings provide insights into the structural mechanism of DmDcr-2 and its cofactors during siRNA processing.


Asunto(s)
Proteínas de Drosophila , Drosophila melanogaster , Animales , ADN Helicasas , Proteínas de Drosophila/genética , Interferencia de ARN , ARN Bicatenario , ARN Interferente Pequeño , Proteínas de Unión al ARN
5.
Nucleic Acids Res ; 51(17): 9475-9490, 2023 09 22.
Artículo en Inglés | MEDLINE | ID: mdl-37587714

RESUMEN

African swine fever virus (ASFV) is highly contagious and can cause lethal disease in pigs. Although it has been extensively studied in the past, no vaccine or other useful treatment against ASFV is available. The genome of ASFV encodes more than 170 proteins, but the structures and functions for the majority of the proteins remain elusive, which hindered our understanding on the life cycle of ASFV and the development of ASFV-specific inhibitors. Here, we report the structural and biochemical studies of the highly conserved C962R protein of ASFV, showing that C962R is a multidomain protein. The N-terminal AEP domain is responsible for the DNA polymerization activity, whereas the DNA unwinding activity is catalyzed by the central SF3 helicase domain. The middle PriCT2 and D5_N domains and the C-terminal Tail domain all contribute to the DNA unwinding activity of C962R. C962R preferentially works on forked DNA, and likely functions in Base-excision repair (BER) or other repair pathway in ASFV. Although it is not essential for the replication of ASFV, C962R can serve as a model and provide mechanistic insight into the replicative primase proteins from many other species, such as nitratiruptor phage NrS-1, vaccinia virus (VACV) and other viruses.


Asunto(s)
Virus de la Fiebre Porcina Africana , Proteínas Virales , Animales , Fiebre Porcina Africana/virología , Virus de la Fiebre Porcina Africana/enzimología , Porcinos , Proteínas Virales/química , Proteínas Virales/metabolismo , ADN-Topoisomerasas de Tipo I/química , Replicación del ADN
6.
Nanoscale ; 15(24): 10223-10231, 2023 Jun 23.
Artículo en Inglés | MEDLINE | ID: mdl-37272869

RESUMEN

Based on the charge-polarity control, a novel anti-ambipolar heterotransistor is proposed based on a special In2Se3&WSe2 van der Waals heterostructure. Unlike traditional logic transistors, our anti-ambipolar heterotransistor can treat an optical signal as an input to change its operating state, that is, with the switching of the optical signal, it shows a reversible polarity change between anti-ambipolar and P-type. Moreover, with the increase of laser power density from 0 to 4.4 mW cm-2, the current value corresponding to the anti-ambipolar peak of the device (Ipeak) shifts from 0 to 4.3 nA, and the voltage value corresponding to the anti-ambipolar peak of the device (Vpeak) can shift from -7 to -5.67 V. These phenomena demonstrate that the charge neutrality point of the anti-ambipolar heterotransistor can be selectively varied with the change of laser power density. In addition, the aforementioned device possesses a high Ion/Ioff ratio of about 104 (405 nm, 4.4 mW cm-2) at 0 V. These properties indicate that our unique anti-ambipolar In2Se3&WSe2 heterotransistor can be employed in photo-triggered inverters for future optoelectronic circuits, and it has great potential to improve the integration of overall chip circuits by implementing optoelectronic logic functions in a unit.

7.
Artículo en Inglés | MEDLINE | ID: mdl-36315112

RESUMEN

Lead-free perovskite has attracted great attention in realizing high-performance optoelectronic devices due to their excellent atmospheric stability and nontoxic characteristics. Although a pronounced ion migration effect has been observed in this new class of materials, its potential in enhancing the overall device performance is yet to be fully explored. In this work, we studied the effect of ion migrations on the carrier transport behavior and found that the recoverable migration process can contribute to enhancing the on/off ratio in a lead-free CsCu2I3 single-crystal microrod-based photodetector. In detail, we synthesized CsCu2I3 single-crystal microrods via an in-plane self-assembly supersaturated crystallization approach. These microrods with well-defined morphologies were then used to construct ultraviolet (UV)-band photodetectors, which outperform most reported lead-free perovskite photodetectors based on individual single crystals. Simultaneously, ion migration can result in asymmetric band bending in the two-terminal device, as confirmed by surface potential profiling with Kelvin probe force microscopy (KPFM). Such an effect can be harnessed to increase the on/off ratio by almost an order of magnitude. Furthermore, the lead-free CsCu2I3 single crystal exhibits excellent thermal and air stabilities. These findings demonstrate that the CsCu2I3 single-crystal microrods can be used in stable and efficient photodetection, and the ion migration effect can potentially be utilized for improving the optoelectronic performance of lead-free devices.

8.
Cell Res ; 32(11): 982-994, 2022 11.
Artículo en Inglés | MEDLINE | ID: mdl-36167981

RESUMEN

N6-methyladenosine (m6A) is the most abundant ribonucleotide modification among eukaryotic messenger RNAs. The m6A "writer" consists of the catalytic subunit m6A-METTL complex (MAC) and the regulatory subunit m6A-METTL-associated complex (MACOM), the latter being essential for enzymatic activity. Here, we report the cryo-electron microscopy (cryo-EM) structures of MACOM at a 3.0-Å resolution, uncovering that WTAP and VIRMA form the core structure of MACOM and that ZC3H13 stretches the conformation by binding VIRMA. Furthermore, the 4.4-Å resolution cryo-EM map of the MACOM-MAC complex, combined with crosslinking mass spectrometry and GST pull-down analysis, elucidates a plausible model of the m6A writer complex, in which MACOM binds to MAC mainly through WTAP and METTL3 interactions. In combination with in vitro RNA substrate binding and m6A methyltransferase activity assays, our results illustrate the molecular basis of how MACOM assembles and interacts with MAC to form an active m6A writer complex.


Asunto(s)
Metiltransferasas , Humanos , Microscopía por Crioelectrón , ARN Mensajero/metabolismo , Metiltransferasas/metabolismo
9.
Nanomaterials (Basel) ; 12(17)2022 Sep 04.
Artículo en Inglés | MEDLINE | ID: mdl-36080110

RESUMEN

In the progress of nonlinear optics, multiphoton absorption (MPA) upconversion lasing enables many vital applications in bioimaging, three-dimensional optical data storage, and photodynamic therapy. Here, efficient four-photon absorption upconversion lasing from the ZnO/ZnMgO multiple quantum wells (MQWs) at room temperature is realized. Moreover, the MPA upconversion lasing and third-harmonic generation peak generated in the MQWs under the excitation of a femtosecond (fs) laser pulse were observed concurrently, and the essential differences between each other were studied comprehensively. Compared with the ZnO film, the upconversion lasing peak of the ZnO/ZnMgO MQWs exhibits a clear blue shift. In addition, the four-photon absorption upconversion photoluminescence (PL) intensity was enhanced in the MQWs/Au nanoparticles (NPs) by the metal-localized surface plasmons (LSPs). The work paves the way for short-wavelength lasers by taking advantage of the high stability and large exciton binding energy of the MQWs' structures.

10.
Nature ; 607(7918): 399-406, 2022 07.
Artículo en Inglés | MEDLINE | ID: mdl-35768513

RESUMEN

Small interfering RNAs (siRNAs) are the key components for RNA interference (RNAi), a conserved RNA-silencing mechanism in many eukaryotes1,2. In Drosophila, an RNase III enzyme Dicer-2 (Dcr-2), aided by its cofactor Loquacious-PD (Loqs-PD), has an important role in generating 21 bp siRNA duplexes from long double-stranded RNAs (dsRNAs)3,4. ATP hydrolysis by the helicase domain of Dcr-2 is critical to the successful processing of a long dsRNA into consecutive siRNA duplexes5,6. Here we report the cryo-electron microscopy structures of Dcr-2-Loqs-PD in the apo state and in multiple states in which it is processing a 50 bp dsRNA substrate. The structures elucidated interactions between Dcr-2 and Loqs-PD, and substantial conformational changes of Dcr-2 during a dsRNA-processing cycle. The N-terminal helicase and domain of unknown function 283 (DUF283) domains undergo conformational changes after initial dsRNA binding, forming an ATP-binding pocket and a 5'-phosphate-binding pocket. The overall conformation of Dcr-2-Loqs-PD is relatively rigid during translocating along the dsRNA in the presence of ATP, whereas the interactions between the DUF283 and RIIIDb domains prevent non-specific cleavage during translocation by blocking the access of dsRNA to the RNase active centre. Additional ATP-dependent conformational changes are required to form an active dicing state and precisely cleave the dsRNA into a 21 bp siRNA duplex as confirmed by the structure in the post-dicing state. Collectively, this study revealed the molecular mechanism for the full cycle of ATP-dependent dsRNA processing by Dcr-2-Loqs-PD.


Asunto(s)
Microscopía por Crioelectrón , Proteínas de Drosophila , Drosophila melanogaster , ARN Helicasas , ARN Bicatenario , ARN Interferente Pequeño , Proteínas de Unión al ARN , Ribonucleasa III , Adenosina Trifosfato , Animales , Sitios de Unión , Proteínas de Drosophila/química , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/ultraestructura , Fosfatos/metabolismo , Conformación Proteica , ARN Helicasas/química , ARN Helicasas/metabolismo , ARN Helicasas/ultraestructura , ARN Bicatenario/química , ARN Bicatenario/metabolismo , ARN Bicatenario/ultraestructura , ARN Interferente Pequeño/química , ARN Interferente Pequeño/metabolismo , ARN Interferente Pequeño/ultraestructura , Proteínas de Unión al ARN/química , Proteínas de Unión al ARN/metabolismo , Proteínas de Unión al ARN/ultraestructura , Ribonucleasa III/química , Ribonucleasa III/metabolismo , Ribonucleasa III/ultraestructura
11.
Nanoscale ; 14(20): 7589-7595, 2022 May 26.
Artículo en Inglés | MEDLINE | ID: mdl-35514279

RESUMEN

Nonlinear multiphoton absorption (MPA) upconversion lasers have critical applications in fluorescence imaging probes and biological photonics. Here, we report the realization of ultralow-threshold six-photon-excited upconversion lasing through cavity quantum electrodynamics effects in a plasmonic microcavity. The value of the Purcell factor (Fp) in hybrid whisper-gallery mode (WGM) is enhanced five-fold relative to a bare microwire (MW), which enhances the nonlinear light-matter interactions dramatically. Compared with a MW, the threshold of six-photon upconversion WGM lasing is reduced by one order magnitude due to plasmonic enhancement effects. In addition, the temperature and polarization characteristics of upconversion lasing via a plasmonic-WGM approach show a distinct evolution, different from a bare MW. This work paves the way for extreme nonlinear optics, taking advantage of the processability and high Purcell factor of plasmonic microcavities.

12.
ACS Appl Mater Interfaces ; 14(18): 21383-21391, 2022 May 11.
Artículo en Inglés | MEDLINE | ID: mdl-35482007

RESUMEN

Recently, identifying promising new two-dimensional (2D) materials with low-symmetry structures has aroused great interest for developing monolithic polarization-sensitive photodetectors with small volume. Here, after comprehensive research of the in-plane anisotropic structure and electronic and optoelectronic properties of layered γ-InSe, a superior responsivity polarization-sensitive photodetector based on multilayer γ-InSe is constructed by a facile method. Notably, the conductance and carrier mobility of the device along the armchair direction are 11.8 and 2.35 times larger than those along the zigzag direction, respectively. Benefitting from the high efficiency of light absorption and excellent carrier mobility (221 cm2 V-1 s-1) of our multilayered γ-InSe along the armchair direction, the device exhibits a superior responsivity of 127 A/W and an external quantum efficiency (EQE) of 104%. Especially, the highest responsivity along the armchair direction of our γ-InSe polarization-sensitive photodetectors can reach as high as 78.5 A/W under polarized light. This value is much higher than those of other devices even under unpolarized light. This work not only provides an insight into the in-plane anisotropic properties of 2D layered γ-InSe but also proposes a stable and environmentally friendly candidate for anisotropic optoelectronic applications.

13.
Nat Commun ; 13(1): 2290, 2022 04 28.
Artículo en Inglés | MEDLINE | ID: mdl-35484139

RESUMEN

Besides the canonical RNA-based RNase P, pre-tRNA 5'-end processing can also be catalyzed by protein-only RNase P (PRORP). To date, various PRORPs have been discovered, but the basis underlying substrate binding and cleavage by HARPs (homolog of Aquifex RNase P) remains elusive. Here, we report structural and biochemical studies of HARPs. Comparison of the apo- and pre-tRNA-complexed structures showed that HARP is able to undergo large conformational changes that facilitate pre-tRNA binding and catalytic site formation. Planctomycetes bacterium HARP exists as dimer in vitro, but gel filtration and electron microscopy analysis confirmed that HARPs from Thermococcus celer, Thermocrinis minervae and Thermocrinis ruber can assemble into larger oligomers. Structural analysis, mutagenesis and in vitro biochemical studies all supported one cooperative pre-tRNA processing mode, in which one HARP dimer binds pre-tRNA at the elbow region whereas 5'-end removal is catalyzed by the partner dimer. Our studies significantly advance our understanding on pre-tRNA processing by PRORPs.


Asunto(s)
Precursores del ARN , Ribonucleasa P , Dominio Catalítico , Precursores del ARN/metabolismo , Procesamiento Postranscripcional del ARN , ARN de Transferencia/metabolismo , Ribonucleasa P/metabolismo
14.
ACS Chem Biol ; 17(4): 854-863, 2022 04 15.
Artículo en Inglés | MEDLINE | ID: mdl-35294178

RESUMEN

Development of tools for precise manipulation of cellular mRNA m6A methylation at the base level is highly required. Here, we report an RNA-guided RNA modification strategy using a fusion protein containing deactivated nuclease Cas13b and m6A methyltransferase METTL14, namely, dCas13b-M14, which is designedly positioned in the cytoplasm. dCas13b-M14 naturally heterodimerizes with endogenous METTL3 to form a catalytic complex to methylate specific cytoplasmic mRNA under a guide RNA (gRNA). We developed assays to screen and validate the guiding specificity of varied gRNAs at single-base resolution. With an optimum combination of dCas13b-M14 and gRNAs inside cells, we have successfully tuned methylation levels of several selected mRNA m6A sites. The off-target effect was evaluated by whole transcriptome m6A sequencing, and a very minor perturbation on the methylome was revealed. Finally, we successfully utilized the editing tool to achieve de novo methylations on five selected mRNA sites. Together, this study paves the way for studying position-dependent roles of m6A methylation in a particular transcript.


Asunto(s)
Metiltransferasas , ARN , Metilación , Metiltransferasas/genética , Metiltransferasas/metabolismo , ARN/metabolismo , ARN Guía de Kinetoplastida/genética , ARN Mensajero/genética , ARN Mensajero/metabolismo
15.
J Phys Chem Lett ; 13(3): 726-732, 2022 Jan 27.
Artículo en Inglés | MEDLINE | ID: mdl-35025526

RESUMEN

Polymeric carbon nitride (PCN) shows great potential applications in the areas of sustainable energy (photocatalysis and photoelectric conversion, as well as other important catalytic reactions), biosensing, biomedicine, devices, and more, but efficient phosphorescence is very scarce because of the lack of an effective synthetic method and an unsettled phosphorescent mechanism. Herein, we report a strategy to promote efficient phosphorescence to activate triplet exciton release by introduction of S and N elements. PCN could be synthesized by thiourea or urea (named S,N-PCN and N-PCN, respectively) at a relatively low reaction temperature (260 °C). S,N-PCN exhibits phosphorescence quantum yield (4.15%) higher than that (0.41%) for N-PCN. The introduction of C=S and C≡N groups in S,N-PCN networks could boost the intersystem crossing (ISC), leading to small singlet-triplet energy (ΔEST) up to more triplet exciton generation. Considering the excellent optical stability of PCN, a preliminary application of visible-light-excited PCN in advanced anticounterfeiting is proposed.

16.
ACS Appl Mater Interfaces ; 13(41): 49058-49065, 2021 Oct 20.
Artículo en Inglés | MEDLINE | ID: mdl-34633792

RESUMEN

With many advantages including superior color saturation and efficiency, quantum dot light-emitting diodes (QLEDs) are considered a promising candidate for the next-generation displays. Emission uniformity over the entire device area is a critical factor to the overall performance and reliability of QLEDs. In this work, we performed a thorough study on the origin of dark spots commonly observed in operating QLEDs and developed a strategy to eliminate these defects. Using advanced cross section fabrication and imaging techniques, we discovered the occurrence of voids in the organic hole transport layer and directly correlated them to the observed emission nonuniformity. Further investigations revealed that these voids are thermal damages induced during the subsequent thermal deposition of other functional layers and can act as leakage paths in the device. By inserting a thermo-tolerant 1,4,5,8,9,11-hexaazatriphenylene-hexacarbonitrile (HATCN) interlayer with an optimized thickness, the thermally induced dark spots can be completely suppressed, leading to a current efficiency increase by 18%. We further demonstrated that such a thermal passivation strategy can work universally for various types of organic layers with low thermal stability. Our findings here provide important guidance in enhancing the performances and reliability of QLEDs and also other sandwich-structured devices via the passivation of heat-sensitive layers.

17.
Mater Horiz ; 8(11): 3113-3123, 2021 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-34545908

RESUMEN

In-plane anisotropic two-dimensional (2D) materials offer great opportunities for developing novel polarization sensitive photodetectors without being in conjunction with filters and polarizers. However, owing to low linear dichroism ratio and insufficient optical absorption of the few layer 2D materials, the comprehensive performance of the present polarization sensitive photodetectors based on 2D materials is still lower than the practical application requirements. In this work, after systematic investigation of the structural, vibrational, and optical anisotropies of layer-structured Te nanosheets, a novel polarization-sensitive self-powered imaging photodetector with high comprehensive performance based on a p-Te/n-MoSe2 van der Waals heterojunction (vdWH) with strong interlayer transition is proposed. Owing to the high rectification ratio (104) of the diode, the device shows excellent photovoltaic characteristics. As examples, the photodetectors exhibited an ultrahigh on/off ratio of 105 at a relatively weak light intensity (4.73 mw cm-2), and the highest responsivity of the device could reach 2106 mA W-1 without any power supply. In particular, benefitting from the excellent dichroism properties of Te nanosheets synthesized in this work, the anisotropic ratio of the photocurrent (Imax/Imin) could reach as high as 16.39 (405 nm, 24.2 mw cm-2). This value obtained under zero bias voltage is much greater than that of present 2D material photodetectors even at a bias voltage. In addition, the highest detectivity is 2.91 × 1013 Jones at a low bias voltage of -0.08 V. This work provides a novel building block for high resolution polarization-sensitive photodetection of weak signals in complex environments.

18.
ACS Appl Mater Interfaces ; 13(32): 38458-38466, 2021 Aug 18.
Artículo en Inglés | MEDLINE | ID: mdl-34348459

RESUMEN

Hybrid organic-inorganic lead halide perovskite microwires are potential building blocks for realizing on-chip integrated optoelectronic devices. However, the length-controllable synthesis of one-dimensional hybrid perovskite microwires has been rarely reported, especially the ones with lengths in the millimeter scale. Herein, methylammonium lead bromide (MAPbBr3) and formamidinium lead bromide (FAPbBr3) micro and milliwires are demonstrated using single-crystal PbBr2 microwires synthesized via template-free solution-phase growth as the lattice framework. Following the PbBr2 template, the as-converted perovskite microwires possess controllable lengths ranging from tens to thousands of micrometers. In addition, Fabry-Perot (FP) lasing was realized in both MAPbBr3 and FAPbBr3 microwires, attesting to their excellent crystal quality and the efficient optical confinement of the natural cavity. These unique properties result in the first demonstration of FP perovskite microwire lasers with submillimeter lengths. More interestingly, the microwire lasers show excellent photostability under repetitive pulsed laser excitation for over 8 × 106 cycles. Such findings demonstrate that the solution-converted hybrid lead bromide microwires have excellent optoelectronic performances promising for practical applications, and the size controllability indicates that this novel fabrication process may be feasible for large-scale industrial production.

19.
PLoS Biol ; 19(4): e3001231, 2021 04.
Artículo en Inglés | MEDLINE | ID: mdl-33905418

RESUMEN

MgtE is a Mg2+ channel conserved in organisms ranging from prokaryotes to eukaryotes, including humans, and plays an important role in Mg2+ homeostasis. The previously determined MgtE structures in the Mg2+-bound, closed-state, and structure-based functional analyses of MgtE revealed that the binding of Mg2+ ions to the MgtE cytoplasmic domain induces channel inactivation to maintain Mg2+ homeostasis. There are no structures of the transmembrane (TM) domain for MgtE in Mg2+-free conditions, and the pore-opening mechanism has thus remained unclear. Here, we determined the cryo-electron microscopy (cryo-EM) structure of the MgtE-Fab complex in the absence of Mg2+ ions. The Mg2+-free MgtE TM domain structure and its comparison with the Mg2+-bound, closed-state structure, together with functional analyses, showed the Mg2+-dependent pore opening of MgtE on the cytoplasmic side and revealed the kink motions of the TM2 and TM5 helices at the glycine residues, which are important for channel activity. Overall, our work provides structure-based mechanistic insights into the channel gating of MgtE.


Asunto(s)
Antiportadores/química , Proteínas Bacterianas/química , Activación del Canal Iónico/fisiología , Antiportadores/metabolismo , Proteínas Bacterianas/metabolismo , Sitios de Unión/efectos de los fármacos , Transporte Biológico , Microscopía por Crioelectrón , Cristalografía por Rayos X , Citoplasma/metabolismo , Activación del Canal Iónico/efectos de los fármacos , Cinética , Magnesio/metabolismo , Magnesio/farmacología , Modelos Moleculares , Dominios Proteicos/efectos de los fármacos , Dominios Proteicos/fisiología , Estructura Cuaternaria de Proteína , Estructura Secundaria de Proteína , Thermus thermophilus/metabolismo
20.
Nucleic Acids Res ; 49(1): 568-583, 2021 Jan 11.
Artículo en Inglés | MEDLINE | ID: mdl-33332555

RESUMEN

Infection with kinetoplastid parasites, including Trypanosoma brucei (T. brucei), Trypanosoma cruzi (T. cruzi) and Leishmania can cause serious disease in humans. Like other kinetoplastid species, mRNAs of these disease-causing parasites must undergo posttranscriptional editing in order to be functional. mRNA editing is directed by gRNAs, a large group of small RNAs. Similar to mRNAs, gRNAs are also precisely regulated. In T. brucei, overexpression of RNase D ribonuclease (TbRND) leads to substantial reduction in the total gRNA population and subsequent inhibition of mRNA editing. However, the mechanisms regulating gRNA binding and cleavage by TbRND are not well defined. Here, we report a thorough structural study of TbRND. Besides Apo- and NMP-bound structures, we also solved one TbRND structure in complexed with single-stranded RNA. In combination with mutagenesis and in vitro cleavage assays, our structures indicated that TbRND follows the conserved two-cation-assisted mechanism in catalysis. TbRND is a unique RND member, as it contains a ZFD domain at its C-terminus. In addition to T. brucei, our studies also advanced our understanding on the potential gRNA degradation pathway in T. cruzi, Leishmania, as well for as other disease-associated parasites expressing ZFD-containing RNDs.


Asunto(s)
Proteínas Protozoarias/química , Estabilidad del ARN/fisiología , ARN Guía de Kinetoplastida/metabolismo , ARN Protozoario/metabolismo , Ribonucleasa III/química , Trypanosoma brucei brucei/enzimología , Secuencia de Aminoácidos , Secuencia de Bases , Cristalografía por Rayos X , Regulación de la Expresión Génica , Modelos Moleculares , Conformación de Ácido Nucleico , Conformación Proteica , Dominios Proteicos , Proteínas Protozoarias/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Ribonucleasa III/metabolismo , Relación Estructura-Actividad , Especificidad por Sustrato , Dedos de Zinc
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