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1.
Plant Cell ; 36(1): 194-212, 2023 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-37804098

RESUMO

In plant leaves, starch is composed of glucan polymers that accumulate in chloroplasts as the products of photosynthesis during the day; starch is mobilized at night to continuously provide sugars to sustain plant growth and development. Efficient starch degradation requires the involvement of several enzymes, including ß-amylase and glucan phosphatase. However, how these enzymes cooperate remains largely unclear. Here, we show that the glucan phosphatase LIKE SEX FOUR 1 (LSF1) interacts with plastid NAD-dependent malate dehydrogenase (MDH) to recruit ß-amylase (BAM1), thus reconstituting the BAM1-LSF1-MDH complex. The starch hydrolysis activity of BAM1 drastically increased in the presence of LSF1-MDH in vitro. We determined the structure of the BAM1-LSF1-MDH complex by a combination of cryo-electron microscopy, crosslinking mass spectrometry, and molecular docking. The starch-binding domain of the dual-specificity phosphatase and carbohydrate-binding module of LSF1 was docked in proximity to BAM1, thus facilitating BAM1 access to and hydrolysis of the polyglucans of starch, thus revealing the molecular mechanism by which the LSF1-MDH complex improves the starch degradation activity of BAM1. Moreover, LSF1 is phosphatase inactive, and the enzymatic activity of MDH was dispensable for starch degradation, suggesting nonenzymatic scaffold functions for LSF1-MDH in starch degradation. These findings provide important insights into the precise regulation of starch degradation.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , beta-Amilase , Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Malato Desidrogenase/metabolismo , beta-Amilase/metabolismo , Simulação de Acoplamento Molecular , Microscopia Crioeletrônica , Amido/metabolismo , Glucanos/metabolismo , Monoéster Fosfórico Hidrolases/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo
2.
BMC Plant Biol ; 24(1): 683, 2024 Jul 18.
Artigo em Inglês | MEDLINE | ID: mdl-39020306

RESUMO

Campanumoea javanica Bl. (CJ) traditionally used in Southwestern China, is now widely consumed as a health food across the nation. Due to its similar efficacy to Codonopsis Radix (CR) and their shared botanical family, CJ is often used as a substitute for CR. According to the Chinese Pharmacopoeia, Codonopsis pilosula var. modesta (Nannf.) L.T. Shen (CPM), Codonopsis pilosula (Franch.) Nannf. (CP), and Codonopsis tangshen Oliv. (CT) are the primary sources of CR. However, details on the differences in composition, effectiveness, and compositional between CJ and CR are still limited. Besides, there is little evidence to support the application of CJ as a drug. In this study, we employed widely targeted metabolomics, network pharmacology analysis, and molecular docking to explore the disparities in metabolite profiles between CJ and CR and to predict the pharmacological mechanisms of the dominant differential metabolites of CJ and their potential medicinal applications. The widely targeted metabolomics results indicated that 1,076, 1,102, 1,102, and 1,093 compounds, most phenolic acids, lipids, amino acids, and flavonoids, were characterized in CJ, CPM, CP, and CT, respectively. There were an average of 1061 shared compounds in CJ and CRs, with 95.07% similarity in metabolic profiles. Most of the metabolites in CJ were previously unreported. Twelve of the seventeen dominant metabolites found in CJ were directly associated with treating cancer and lactation, similar to the traditional medicinal efficacy. The molecular docking results showed that the dominant metabolites of CJ had good docking activity with the core targets PIK3R1, PIK3CA, ESR1, HSP90AA1, EGFR, and AKT1. This study provides a scientific basis for understanding the similarities and differences between CJ and CR at the metabolome level, offering a theoretical foundation for developing innovative medications from CJ. Additionally, it significantly enhances the metabolite databases for both CJ and CR.


Assuntos
Codonopsis , Metabolômica , Farmacologia em Rede , Codonopsis/química , Codonopsis/metabolismo , Simulação de Acoplamento Molecular , Medicamentos de Ervas Chinesas/metabolismo , Medicamentos de Ervas Chinesas/farmacologia , Medicamentos de Ervas Chinesas/química , Raízes de Plantas/metabolismo , Extratos Vegetais/farmacologia , Extratos Vegetais/química , Extratos Vegetais/metabolismo
3.
Nano Lett ; 23(16): 7576-7583, 2023 Aug 23.
Artigo em Inglês | MEDLINE | ID: mdl-37535801

RESUMO

Using in situ atomic-resolution scanning transmission electron microscopy, atomic movements and rearrangements associated with diffusive solid to solid phase transformations in the Pt-Sn system are captured to reveal details of the underlying atomistic mechanisms that drive these transformations. In the PtSn4 to PtSn2 phase transformation, a periodic superlattice substructure and a unique intermediate structure precede the nucleation and growth of the PtSn2 phase. At the atomic level, all stages of the transformation are templated by the anisotropic crystal structure of the parent PtSn4 phase. In the case of the PtSn2 to Pt2Sn3 transformation, the anisotropy in the structure of product Pt2Sn3 dictates the path of transformation. Analysis of atomic configurations at the transformation front elucidates the diffusion pathways and lattice distortions required for these phase transformations. Comparison of multiple Pt-Sn phase transformations reveals the structural parameters governing solid to solid phase transformations in this technologically interesting intermetallic system.

4.
Zhongguo Zhong Yao Za Zhi ; 49(11): 3113-3124, 2024 Jun.
Artigo em Zh | MEDLINE | ID: mdl-39041170

RESUMO

This study aims to analyze the current situation of outcome indicators in randomized controlled trial(RCT) of traditional Chinese medicine(TCM) treatment for Alzheimer's disease(AD), so as to provide a reference for establishing a core indicator set in this field. The researchers systematically searched CNKI, Wanfang, VIP, Sino Med, EMbase, PubMed, Medline, and Cochrane Library. Independent screening of literature and extraction of information was conducted according to the inclusion and exclusion criteria. In addition, the Ro B 2. 0 tool was used for bias risk assessment. A total of 78 RCTs were included, involving 6 379 patients,with 122 kinds of outcome indicators. According to functional attributes, the outcome indicators could be categorized into seven groups:TCM diseases(3 kinds, 13 times), symptoms and signs(26 kinds, 196 times), physical and chemical tests(68 kinds, 149 times),qua-lity of life(1 kind, 2 times), long-term prognosis(2 kinds, 2 times), economic evaluation(0 kind), safety events(21 kinds,194 times), and other indicators(1 kind, 1 time). The results show that the literature evaluation of RCTs of TCM treatment for AD is generally risky, and there are some problems in the selection of outcome indicators, such as lack of TCM characteristics, insignificant distinction between primary and secondary outcome indicators, lack of long-term prognosis and economic evaluation indicators, and non-standard safety event reports. It is suggested that future researchers should establish a core indicator set for AD that highlights the characteristics of TCM and then work to improve the quality of clinical trials.


Assuntos
Doença de Alzheimer , Medicamentos de Ervas Chinesas , Medicina Tradicional Chinesa , Ensaios Clínicos Controlados Aleatórios como Assunto , Doença de Alzheimer/tratamento farmacológico , Humanos , Medicamentos de Ervas Chinesas/uso terapêutico , Resultado do Tratamento , Idoso
5.
Plant Biotechnol J ; 21(9): 1827-1838, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37353991

RESUMO

Bacillus thuringiensis (Bt)-secreted crystal (Cry) toxins form oligomeric pores in host cell membranes and are a common element in generating insect-resistant transgenic crops. Although Cry toxin function has been well documented, cellular defences against pore-formation have not been as well developed. Elucidation of the processes underlying this defence, however, could contribute to the development of enhanced Bt crops. Here, we demonstrate that Cry1Ca-mediated downregulation of microRNA-7322-5p (miR-7322-5p), which binds to the 3' untranslated region of p38, negatively regulates the susceptibility of Chilo suppressalis to Cry1Ca. Moreover, Cry1Ca exposure enhanced phosphorylation of Hsp19, and hsp19 downregulation increased susceptibility to Cry1Ca. Further, Hsp19 phosphorylation occurs downstream of p38, and pull-down assays confirmed the interactions between Hsp19 and Cry1Ca, suggesting that activation of Hsp19 by the miR-7322-5p/p38/Hsp19 pathway promotes Cry1Ca sequestration. To assess the efficacy of targeting this pathway in planta, double-stranded RNA (dsRNA) targeting C. suppressalis p38 (dsp38) was introduced into a previously generated cry1Ca-expressing rice line (1CH1-2) to yield a single-copy cry1Ca/dsp38 rice line (p38-rice). Feeding on this rice line triggered a significant reduction in C. suppressalis p38 expression and the line was more resistant to C. suppressalis than 1CH1-2 in both short term (7-day) and continuous feeding bioassays as well as field trials. These findings provide new insights into invertebrate epithelium cellular defences and demonstrate a potential new pyramiding strategy for Bt crops.


Assuntos
Bacillus thuringiensis , MicroRNAs , Mariposas , Oryza , Animais , Oryza/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Larva/genética , Controle Biológico de Vetores , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Mariposas/fisiologia , Endotoxinas/genética , Endotoxinas/metabolismo , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/metabolismo , Bacillus thuringiensis/genética , Bacillus thuringiensis/metabolismo
6.
Nano Lett ; 22(2): 622-629, 2022 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-34982564

RESUMO

Perpendicular magnetic tunnel junctions (p-MTJs) switched utilizing bipolar electric fields have extensive applications in energy-efficient memory and logic devices. Voltage-controlled magnetic anisotropy linearly lowers the energy barrier of the ferromagnetic layer via the electric field effect and efficiently switches p-MTJs only with a unipolar behavior. Here, we demonstrate a bipolar electric field effect switching of 100 nm p-MTJs with a synthetic antiferromagnetic free layer through voltage-controlled exchange coupling (VCEC). The switching current density, ∼1.1 × 105 A/cm2, is 1 order of magnitude lower than that of the best-reported spin-transfer torque devices. Theoretical results suggest that the electric field induces a ferromagnetic-antiferromagnetic exchange coupling transition of the synthetic antiferromagnetic free layer and generates a fieldlike interlayer exchange coupling torque, which causes the bidirectional magnetization switching of p-MTJs. These results could eliminate the major obstacle in the development of spin memory devices beyond their embedded applications.

7.
Plant J ; 106(3): 630-648, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33547692

RESUMO

The mechanism that coordinates cell growth and cell cycle progression remains poorly understood; in particular, whether the cell cycle and cell wall biosynthesis are coordinated remains unclear. Recently, cell wall biosynthesis and cell cycle progression were reported to respond to wounding. Nonetheless, no genes are reported to synchronize the biosynthesis of the cell wall and the cell cycle. Here, we report that wounding induces the expression of genes associated with cell wall biosynthesis and the cell cycle, and that two genes, AtMYB46 in Arabidopsis thaliana and RrMYB18 in Rosa rugosa, are induced by wounding. We found that AtMYB46 and RrMYB18 promote the biosynthesis of the cell wall by upregulating the expression of cell wall-associated genes, and that both of them also upregulate the expression of a battery of genes associated with cell cycle progression. Ultimately, this response leads to the development of curled leaves of reduced size. We also found that the coordination of cell wall biosynthesis and cell cycle progression by AtMYB46 and RrMYB18 is evolutionarily conservative in multiple species. In accordance with wounding promoting cell regeneration by regulating the cell cycle, these findings also provide novel insight into the coordination between cell growth and cell cycle progression and a method for producing miniature plants.


Assuntos
Arabidopsis/metabolismo , Ciclo Celular , Parede Celular/metabolismo , Rosa/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas , Genes de Plantas/genética , Genes de Plantas/fisiologia , Filogenia , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/fisiologia , Rosa/genética , Fatores de Transcrição/genética , Fatores de Transcrição/fisiologia , Transcriptoma
8.
Nature ; 534(7608): 575-8, 2016 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-27281194

RESUMO

Chemical modifications of RNA have essential roles in a vast range of cellular processes. N(6)-methyladenosine (m(6)A) is an abundant internal modification in messenger RNA and long non-coding RNA that can be dynamically added and removed by RNA methyltransferases (MTases) and demethylases, respectively. An MTase complex comprising methyltransferase-like 3 (METTL3) and methyltransferase-like 14 (METTL14) efficiently catalyses methyl group transfer. In contrast to the well-studied DNA MTase, the exact roles of these two RNA MTases in the complex remain to be elucidated. Here we report the crystal structures of the METTL3-METTL14 heterodimer with MTase domains in the ligand-free, S-adenosyl methionine (AdoMet)-bound and S-adenosyl homocysteine (AdoHcy)-bound states, with resolutions of 1.9, 1.71 and 1.61 Å, respectively. Both METTL3 and METTL14 adopt a class I MTase fold and they interact with each other via an extensive hydrogen bonding network, generating a positively charged groove. Notably, AdoMet was observed in only the METTL3 pocket and not in METTL14. Combined with biochemical analysis, these results suggest that in the m(6)A MTase complex, METTL3 primarily functions as the catalytic core, while METTL14 serves as an RNA-binding platform, reminiscent of the target recognition domain of DNA N(6)-adenine MTase. This structural information provides an important framework for the functional investigation of m(6)A.


Assuntos
Adenosina/análogos & derivados , Biocatálise , Metiltransferases/química , Metiltransferases/metabolismo , Adenosina/metabolismo , Sítios de Ligação , Domínio Catalítico , Humanos , Ligação de Hidrogênio , Ligantes , Metilação , Modelos Biológicos , Modelos Moleculares , Estrutura Quaternária de Proteína , Subunidades Proteicas/química , Subunidades Proteicas/metabolismo , RNA/química , RNA/metabolismo , Relação Estrutura-Atividade
9.
Environ Res ; 212(Pt D): 113391, 2022 09.
Artigo em Inglês | MEDLINE | ID: mdl-35597293

RESUMO

Taihu Lake is the most important drinking water source of the major cities in the Yangtze River Delta. The pollution of endocrine disruptors (EDCs)in Taihu Lake has been increasing recently, the accurate determination is an important guide for predicting its health risks and developing appropriate controls. Monitoring organic pollutants in water using the diffusive gradient in thin film technique (DGT) has attracted much attention due to more accuracy and convenience than the grab sampling methods. In this study, a novel cyclodextrin polymer (CDP) synthesized by the simple and green method in water was taken as an adsorbent for the binding gel. Four endocrine-disrupting chemicals (EDCs), bisphenol A (BPA), 17α-ethinylestradiol (EE2), 17ß-estradiol (E2), and estriol (E3), were taken as models to determine the diffusion coefficients (4.68 × 10-6, 3.38 × 10-6, 3.34 × 10-6 and 4.31 × 10-6 cm2/s) and to test the performance of DGT, such as adsorption capacity and deployment time (1-5 day). The assembled CDP-DGT was adopted to determine four EDCs in a simulated water environment (3-9 of pH, 0.001-0.5 M of ionic strength (IS), and dissolved organic matter (DOM) of 0-20 mg/L). The ability of CDP-DGT sampling was verified in the Jiuxiang River and was carried out for a large-scale field application of in situ sampling EDCs in Taihu Lake basin. The results show that the total EDCs concentration range and the estradiol equivalent concentrations (EEQ) in Taihu Lake and its main rivers are 2.78 ng/L to 11.08 ng/L and 2.62 ng/L to 10.91 ng/L, respectively. The risk quotients (RQs) of all sampling sites in the region were greater than 1, indicating that EDCs pose a serious threat to aquatic organisms in the area. Therefore, the monitoring of EDCs in the Taihu Lake basin should be further strengthened.


Assuntos
Disruptores Endócrinos , Poluentes Químicos da Água , Celulose , China , Ciclodextrinas , Disruptores Endócrinos/análise , Monitoramento Ambiental/métodos , Estradiol , Géis , Lagos/química , Medição de Risco , Rios/química , Água , Poluentes Químicos da Água/análise
10.
Pestic Biochem Physiol ; 187: 105183, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36127045

RESUMO

BACKGROUND: In recent years, gene knockdown technology using double-stranded RNA (dsRNA) has been widely used as an environment-friendly pest control strategy, but its instability and limited cellular uptake have limited its overall effect. Studies have shown that the efficiency of single dsRNA can be improved by using various nanomaterials. However, the effect of stacked-dsRNA wrapped by nanomaterial on pests remains unclear. In the present study, both CYP15C1 and C-factor genes were cloned from the midgut of C. suppressalis, and the transcript of C-factor is most highly expressed in heads. Feeding a dsCYP15C1 or dsC-factor - nanomaterial mixture can downregulate the gene expression and significantly increase larval mortality. More importantly, feeding the stacked-dsRNA wrapped by nanomaterial can significantly increase the mortality of C. suppressalis, compared with feeding dsCYP15C1 or dsC-factor - nanomaterial mixture alone. These results showed that CYP15C1 and C-factor could be potential targets for an effective management of C. suppressalis, and we developed a nanoparticle-facilitated stacked-dsRNA strategy in the control of C. suppresallis. Our research provides a theoretical basis for gene function analysis and field pest control, and will promote the application of RNAi technology in the stacked style of pest control.


Assuntos
Mariposas , Nanopartículas , Animais , Larva/genética , Mariposas/genética , Interferência de RNA , RNA de Cadeia Dupla/genética
11.
Molecules ; 27(3)2022 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-35163887

RESUMO

With the internationalization of traditional Chinese medicines (TCMs) and the increasing use of herbal medicines around the world, there are concerns over their safety. In recent years, there have been some sporadic reports of pesticide residues in Chuanxiong Rhizoma (CX), although the lack of systematic and comprehensive analyses of pesticide residues and evaluations of toxicological risks in human health has increased the uncertainty of the potential effects of pesticides exposure in humans. This study aimed to clarify the status of pesticide residues and to determine the health risks of pesticide residues in CX. The findings of this study revealed that 99 batches of CX samples contained pesticide residues ranging from 0.05 to 3013.17 µg/kg. Here, 6-22 kinds of pesticides were detected in each sample. Prometryn, carbendazim, dimethomorph, chlorpyrifos, chlorantraniliprole, pyraclostrobin, and paclobutrazol were the most frequently detected pesticides, with detection rates of 68.69-100%. Insecticides and fungicides accounted for 43.23% and 37.84% of the total pesticides detected, respectively. Here, 86.87% of the pesticide content levels were lower than 50 µg/kg, and a small number of samples contained carbofuran, dimethoate, and isofenphos-methyl exceeding the maximum residue levels (MRLs). A risk assessment based on the hazard quotient/hazard index (HQ/HI) approach revealed that the short-term, long-term, and cumulative risks of pesticide residues in CX are well below the levels that may pose a health risk. Worryingly, six banned pesticides (carbofuran, phorate sulfone, phorate-sulfoxide, isofenphos-methyl, terbufos-sulfone, and terbufoxon sulfoxide) were detected. This study has improved our understanding of the potential exposure risk of pesticide multi-residues in CX. The results of the study will have a positive impact on improving the quality and safety of CX and the development of MRLs for pesticide residues.


Assuntos
Cromatografia Gasosa/métodos , Cromatografia Líquida/métodos , Medicamentos de Ervas Chinesas/análise , Contaminação de Alimentos/análise , Medicina Tradicional Chinesa , Resíduos de Praguicidas/análise , Espectrometria de Massas em Tandem/métodos , Humanos , Medição de Risco
12.
Plant Biotechnol J ; 19(8): 1579-1587, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-33638282

RESUMO

Cellulose is one of the most abundant organic polymers in nature. It contains multiple ß-1,4-glucan chains synthesized by cellulose synthases (CesAs) on the plasma membrane of higher plants. CesA subunits assemble into a pseudo-sixfold symmetric cellulose synthase complex (CSC), known as a 'rosette complex'. The structure of CesA remains enigmatic. Here, we report the cryo-EM structure of the homotrimeric CesA7 from Gossypium hirsutum at 3.5-angstrom resolution. The GhCesA7 homotrimer shows a C3 symmetrical assembly. Each protomer contains seven transmembrane helices (TMs) which form a channel potentially facilitating the release of newly synthesized glucans. The cytoplasmic glycosyltransferase domain (GT domain) of GhCesA7 protrudes from the membrane, and its catalytic pocket is directed towards the TM pore. The homotrimer GhCesA7 is stabilized by the transmembrane helix 7 (TM7) and the plant-conserved region (PCR) domains. It represents the building block of CSCs and facilitates microfibril formation. This structure provides insight into how eukaryotic cellulose synthase assembles and provides a mechanistic basis for the improvement of cotton fibre quality in the future.


Assuntos
Glucosiltransferases , Gossypium , Celulose , Fibra de Algodão , Glucosiltransferases/genética , Gossypium/genética
13.
RNA Biol ; 18(sup1): 244-253, 2021 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-34074215

RESUMO

Various kinds of cap structures, such as m7G, triphosphate groups, NAD and dpCoA, protect the 5' terminus of RNA. The cap structures bond covalently to RNA and affect its stability, translation, and transport. The removal of the caps is mainly executed by Nudix hydrolase family proteins, including Dcp2, RppH and NudC. Numerous efforts have been made to elucidate the mechanism underlying the removal of m7G, triphosphate group, and NAD caps. In contrast, few studies related to the cleavage of the RNA dpCoA cap have been conducted. Here, we report the hydrolytic activity of Escherichia coli NudC towards dpCoA and dpCoA-capped RNA in vitro. We also determined the crystal structure of dimeric NudC in complex with dpCoA at 2.0 Å resolution. Structural analysis revealed that dpCoA is recognized and hydrolysed in a manner similar to NAD. In addition, NudC may also remove other dinucleotide derivative caps of RNA, which comprise the AMP moieties. NudC homologs in Saccharomyces cerevisiae and Arabidopsis thaliana exhibited similar dpCoA decapping (deCoAping) activity. These results together indicate a conserved mechanism underpinning the hydrolysis of dpCoA-capped RNA in both prokaryotes and eukaryotes.


Assuntos
Coenzima A/metabolismo , Proteínas de Escherichia coli/metabolismo , Escherichia coli/metabolismo , Pirofosfatases/metabolismo , Capuzes de RNA/química , RNA Bacteriano/química , Escherichia coli/genética , Proteínas de Escherichia coli/química , Proteínas de Escherichia coli/genética , Conformação Proteica , Pirofosfatases/química , Pirofosfatases/genética , Capuzes de RNA/genética , Capuzes de RNA/metabolismo , RNA Bacteriano/genética , RNA Bacteriano/metabolismo , Nudix Hidrolases
14.
Med Sci Monit ; 27: e934297, 2021 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-34355705

RESUMO

Figure 1 is updated due to the mistake that occurred during the layout process. Reference: Aiqing Nie, Yueyue Xiao, Si Liu, Xiaolei Zhu, Delin Zhang. Sensitivity of Reality Monitoring to Fluency: Evidence from Behavioral Performance and Event-Related Potential (ERP) Old/New Effects. Med Sci Monit. 2019; 25: 9490-9498. 10.12659/MSM.917401.

15.
Nucleic Acids Res ; 47(7): 3728-3738, 2019 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-30753696

RESUMO

Members of the pentatricopeptide repeat (PPR) protein family are sequence-specific RNA-binding proteins that play crucial roles in organelle RNA metabolism. Each PPR protein consists of a tandem array of PPR motifs, each of which aligns to one nucleotide of the RNA target. The di-residues in the PPR motif, which are referred to as the PPR codes, determine nucleotide specificity. Numerous PPR codes are distributed among the vast number of PPR motifs, but the correlation between PPR codes and RNA bases is poorly understood, which hinders target RNA prediction and functional investigation of PPR proteins. To address this issue, we developed a modular assembly method for high-throughput construction of designer PPRs, and by using this method, 62 designer PPR proteins containing various PPR codes were assembled. Then, the correlation between these PPR codes and RNA bases was systematically explored and delineated. Based on this correlation, the web server PPRCODE (http://yinlab.hzau.edu.cn/pprcode) was developed. Our study will not only serve as a platform for facilitating target RNA prediction and functional investigation of the large number of PPR family proteins but also provide an alternative strategy for the assembly of custom PPRs that can potentially be used for plant organelle RNA manipulation.


Assuntos
Proteínas de Arabidopsis/genética , Motivos de Nucleotídeos/genética , Proteínas de Ligação a RNA/genética , RNA/genética , Sequência de Aminoácidos/genética , Arabidopsis/genética , Modelos Moleculares , Organelas/genética
16.
Nucleic Acids Res ; 46(13): 6880-6892, 2018 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-29931175

RESUMO

mRNA decay is an important strategy by which bacteria can rapidly adapt to their ever-changing surroundings. The 5'-terminus state of mRNA determines the velocity of decay of many types of RNA. In Escherichia coli, RNA pyrophosphohydrolase (RppH) is responsible for the removal of the 5'-terminal triphosphate from hundreds of mRNAs and triggers its rapid degradation by ribonucleases. A diaminopimelate epimerase, DapF, can directly interact with RppH and stimulate its hydrolysis activity in vivo and in vitro. However, the molecular mechanism remains to be elucidated. Here, we determined the complex structure of DapF-RppH as a heterotetramer in a 2:2 molar ratio. DapF-bound RppH exhibits an RNA-favorable conformation similar to the RNA-bound state, suggesting that association with DapF promotes and stabilizes RppH in a conformation that facilitates substrate RNA binding and thus stimulates the activity of RppH. To our knowledge, this is the first published structure of an RNA-pyrophosphohydrolysis complex in bacteria. Our study provides a framework for further investigation of the potential regulators involved in the RNA-pyrophosphohydrolysis process in prokaryotes.


Assuntos
Hidrolases Anidrido Ácido/química , Isomerases de Aminoácido/química , Proteínas de Escherichia coli/química , Hidrolases Anidrido Ácido/metabolismo , Isomerases de Aminoácido/metabolismo , Cristalografia por Raios X , Proteínas de Escherichia coli/metabolismo , Modelos Moleculares , Ligação Proteica , Conformação Proteica , Multimerização Proteica , RNA/química , RNA/metabolismo
17.
Artigo em Inglês | MEDLINE | ID: mdl-33654328

RESUMO

Materials such as L10 Fe-based alloys with perpendicular magnetic anisotropy derived from crystal structure have the potential to deliver higher thermal stability of magnetic memory elements compared to materials whose anisotropy is derived from surfaces and interfaces. A number of processing parameters enable control of the quality and texture of L10 FePd among them, including substrate, deposition temperature, pressure and seed and buffer layer. The angle of inclination between the substrate and the sputtering target can also impact the texture of L10 crystallization of sputtered Fe-Pd and magnetic properties of the derived thin films. This study examines the difference between FePd layers that have been magnetron sputter deposited on Cr(15 nm)/Pt, Ir, or Ru(4 nm)/FePd (8 nm)/Ru(2 nm)/Ta(3 nm) substrate layers at an oblique angle (30° tilt from the sputtering target) versus normal incidence (target facing the substrate). X-ray diffraction, ferromagnetic resonance spectroscopy and vibrating sample magnetometry were used to compare the degree of L10 order and static and dynamic properties of films deposited under both conditions. The films grown using the oblique orientation exhibit a stronger degree of L10 orientation, a larger magnetic anisotropy energy and a lower Gilbert damping, on all three buffer layers.

18.
Mol Cancer ; 18(1): 186, 2019 12 19.
Artigo em Inglês | MEDLINE | ID: mdl-31856849

RESUMO

BACKGROUND: N6-methyladenosine (m6A) modification, the most abundant internal methylation of eukaryotic RNA transcripts, is critically implicated in RNA processing. As the largest known component in the m6A methyltransferase complex, KIAA1429 plays a vital role in m6A methylation. However, its function and mechanism in hepatocellular carcinoma (HCC) remain poorly defined. METHODS: Quantitative PCR, western blot and immunohistochemistry were used to measure the expression of KIAA1429 in HCC. The effects of KIAA1429 on the malignant phenotypes of hepatoma cells were examined in vitro and in vivo. MeRIP-seq, RIP-seq and RNA-seq were performed to identify the target genes of KIAA1429. RESULTS: KIAA1429 was considerably upregulated in HCC tissues. High expression of KIAA1429 was associated with poor prognosis among HCC patients. Silencing KIAA1429 suppressed cell proliferation and metastasis in vitro and in vivo. GATA3 was identified as the direct downstream target of KIAA1429-mediated m6A modification. KIAA1429 induced m6A methylation on the 3' UTR of GATA3 pre-mRNA, leading to the separation of the RNA-binding protein HuR and the degradation of GATA3 pre-mRNA. Strikingly, a long noncoding RNA (lncRNA) GATA3-AS, transcribed from the antisense strand of the GATA3 gene, functioned as a cis-acting element for the preferential interaction of KIAA1429 with GATA3 pre-mRNA. Accordingly, we found that the tumor growth and metastasis driven by KIAA1429 or GATA3-AS were mediated by GATA3. CONCLUSION: Our study proposed a complex KIAA1429-GATA3 regulatory model based on m6A modification and provided insights into the epi-transcriptomic dysregulation in hepatocarcinogenesis and metastasis.


Assuntos
Adenosina/análogos & derivados , Fator de Transcrição GATA3/genética , Neoplasias Hepáticas/genética , Neoplasias Hepáticas/metabolismo , Processamento Pós-Transcricional do RNA , Proteínas de Ligação a RNA/genética , Adenosina/metabolismo , Animais , Linhagem Celular Tumoral , Modelos Animais de Doenças , Progressão da Doença , Fator de Transcrição GATA3/metabolismo , Humanos , Neoplasias Hepáticas/mortalidade , Neoplasias Hepáticas/patologia , Metilação , Camundongos , Modelos Biológicos , Metástase Neoplásica , Prognóstico , RNA Antissenso/genética , RNA Longo não Codificante/genética , Proteínas de Ligação a RNA/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
19.
Nat Mater ; 17(9): 800-807, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30061733

RESUMO

The spin-orbit torque (SOT) that arises from materials with large spin-orbit coupling promises a path for ultralow power and fast magnetic-based storage and computational devices. We investigated the SOT from magnetron-sputtered BixSe(1-x) thin films in BixSe(1-x)/Co20Fe60B20 heterostructures by using d.c. planar Hall and spin-torque ferromagnetic resonance (ST-FMR) methods. Remarkably, the spin torque efficiency (θS) was determined to be as large as 18.62 ± 0.13 and 8.67 ± 1.08 using the d.c. planar Hall and ST-FMR methods, respectively. Moreover, switching of the perpendicular CoFeB multilayers using the SOT from the BixSe(1-x) was observed at room temperature with a low critical magnetization switching current density of 4.3 × 105 A cm-2. Quantum transport simulations using a realistic sp3 tight-binding model suggests that the high SOT in sputtered BixSe(1-x) is due to the quantum confinement effect with a charge-to-spin conversion efficiency that enhances with reduced size and dimensionality. The demonstrated θS, ease of growth of the films on a silicon substrate and successful growth and switching of perpendicular CoFeB multilayers on BixSe(1-x) films provide an avenue for the use of BixSe(1-x) as a spin density generator in SOT-based memory and logic devices.

20.
Plant Physiol ; 177(2): 684-697, 2018 06.
Artigo em Inglês | MEDLINE | ID: mdl-29686056

RESUMO

Chloroplast biogenesis and development are highly complex processes requiring interaction between plastid and nuclear genomic products. Using a high-throughput screen for chloroplast biogenesis suppressors in Arabidopsis (Arabidopsis thaliana), we identified a suppressor of thf1 (sot5) that displays virescent and serrated leaves. Further characterization revealed that sot5 mutants are defective in leaf adaxial and abaxial polarity and act as enhancers of asymmetric leaves2 Map-based cloning identified SOT5 as a gene previously named EMB2279 that encodes a plastid-targeted pentatricopeptide repeat (PPR) protein with 11 PPR motifs. A G-to-A mutation in sot5 leads to a significant decrease in splicing efficiency, generating two additional mRNA variants. As reported previously, the sot5 null mutation is embryo lethal. SOT5 is predicted to bind to specific RNA sequences found in plastid rpl2 and trnK genes, and we found decreased splicing efficiency of the rpl2 and trnK genes in sot5 mutants. Together, our results reveal that the PPR protein SOT5/EMB2279 is required for intron splicing of plastid rpl2 and trnK, providing insights into the role of plastid translation in the coupled development between chloroplasts and leaves.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Proteínas de Cloroplastos/genética , Íntrons , Plastídeos/genética , Proteases Dependentes de ATP/genética , Arabidopsis/citologia , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/metabolismo , Proliferação de Células/genética , Proteínas de Cloroplastos/metabolismo , Cloroplastos/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Membrana/genética , Mutação , Células Vegetais/patologia , Folhas de Planta/citologia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Plantas Geneticamente Modificadas , RNA Ribossômico/genética , RNA Ribossômico/metabolismo
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