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1.
Cell ; 153(5): 1012-24, 2013 May 23.
Article in English | MEDLINE | ID: mdl-23706739

ABSTRACT

Histone acetylation plays critical roles in chromatin remodeling, DNA repair, and epigenetic regulation of gene expression, but the underlying mechanisms are unclear. Proteasomes usually catalyze ATP- and polyubiquitin-dependent proteolysis. Here, we show that the proteasomes containing the activator PA200 catalyze the polyubiquitin-independent degradation of histones. Most proteasomes in mammalian testes ("spermatoproteasomes") contain a spermatid/sperm-specific α subunit α4 s/PSMA8 and/or the catalytic ß subunits of immunoproteasomes in addition to PA200. Deletion of PA200 in mice abolishes acetylation-dependent degradation of somatic core histones during DNA double-strand breaks and delays core histone disappearance in elongated spermatids. Purified PA200 greatly promotes ATP-independent proteasomal degradation of the acetylated core histones, but not polyubiquitinated proteins. Furthermore, acetylation on histones is required for their binding to the bromodomain-like regions in PA200 and its yeast ortholog, Blm10. Thus, PA200/Blm10 specifically targets the core histones for acetylation-mediated degradation by proteasomes, providing mechanisms by which acetylation regulates histone degradation, DNA repair, and spermatogenesis.


Subject(s)
DNA Repair , Histones/metabolism , Nuclear Proteins/metabolism , Proteasome Endopeptidase Complex/metabolism , Spermatogenesis , Testis/metabolism , Acetylation , Amino Acid Sequence , Animals , DNA Breaks, Double-Stranded , Humans , Male , Mice , Molecular Sequence Data , Nuclear Proteins/chemistry , Protein Structure, Tertiary , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Sequence Alignment
2.
EMBO Rep ; 25(2): 770-795, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38182816

ABSTRACT

DExD/H-box helicases are crucial regulators of RNA metabolism and antiviral innate immune responses; however, their role in bacteria-induced inflammation remains unclear. Here, we report that DDX5 interacts with METTL3 and METTL14 to form an m6A writing complex, which adds N6-methyladenosine to transcripts of toll-like receptor (TLR) 2 and TLR4, promoting their decay via YTHDF2-mediated RNA degradation, resulting in reduced expression of TLR2/4. Upon bacterial infection, DDX5 is recruited to Hrd1 at the endoplasmic reticulum in an MyD88-dependent manner and is degraded by the ubiquitin-proteasome pathway. This process disrupts the DDX5 m6A writing complex and halts m6A modification as well as degradation of TLR2/4 mRNAs, thereby promoting the expression of TLR2 and TLR4 and downstream NF-κB activation. The role of DDX5 in regulating inflammation is also validated in vivo, as DDX5- and METTL3-KO mice exhibit enhanced expression of inflammatory cytokines. Our findings show that DDX5 acts as a molecular switch to regulate inflammation during bacterial infection and shed light on mechanisms of quiescent inflammation during homeostasis.


Subject(s)
Adenine , Bacterial Infections , Toll-Like Receptor 2 , Animals , Mice , Adenine/analogs & derivatives , Inflammation/genetics , Methyltransferases/genetics , Toll-Like Receptor 2/genetics , Toll-Like Receptor 4/genetics
3.
Proc Natl Acad Sci U S A ; 120(35): e2304242120, 2023 08 29.
Article in English | MEDLINE | ID: mdl-37607234

ABSTRACT

Zoonotic poxviruses such as mpox virus (MPXV) continue to threaten public health safety since the eradication of smallpox. Vaccinia virus (VACV), the prototypic poxvirus used as the vaccine strain for smallpox eradication, is the best-characterized member of the poxvirus family. VACV encodes a serine protease inhibitor 1 (SPI-1) conserved in all orthopoxviruses, which has been recognized as a host range factor for modified VACV Ankara (MVA), an approved smallpox vaccine and a promising vaccine vector. FAM111A (family with sequence similarity 111 member A), a nuclear protein that regulates host DNA replication, was shown to restrict the replication of a VACV SPI-1 deletion mutant (VACV-ΔSPI-1) in human cells. Nevertheless, the detailed antiviral mechanisms of FAM111A were unresolved. Here, we show that FAM111A is a potent restriction factor for VACV-ΔSPI-1 and MVA. Deletion of FAM111A rescued the replication of MVA and VACV-ΔSPI-1 and overexpression of FAM111A significantly reduced viral DNA replication and virus titers but did not affect viral early gene expression. The antiviral effect of FAM111A necessitated its trypsin-like protease domain and DNA-binding domain but not the PCNA-interacting motif. We further identified that FAM111A translocated into the cytoplasm upon VACV infection by degrading the nuclear pore complex via its protease activity, interacted with VACV DNA-binding protein I3, and promoted I3 degradation through autophagy. Moreover, SPI-1 from VACV, MPXV, or lumpy skin disease virus was able to antagonize FAM111A by prohibiting its nuclear export. Our findings reveal the detailed mechanism by which FAM111A inhibits VACV and provide explanations for the immune evasive function of VACV SPI-1.


Subject(s)
Poxviridae , Smallpox , Vaccinia , Animals , Cattle , Humans , Vaccinia virus/genetics , Serine Proteinase Inhibitors , Viral Proteins/genetics , DNA Replication , Host Specificity , DNA, Viral , Virus Replication , Receptors, Virus
4.
Plant J ; 117(4): 1130-1147, 2024 Feb.
Article in English | MEDLINE | ID: mdl-37967025

ABSTRACT

Flowering is an indicator of plant transformation from vegetative to reproductive growth. miR160 has been shown to have a significant effect on the growth and development of fruits, leaves, and roots of plants or their stress response to environment, but the participation of miR160 in regulating flowering time in plants is unclear. In this study, we found that two FvemiR160s (FvemiR160a/FvemiR160b) mature sequences in strawberry (Fragaria vesca) were consistent. It was displayed that the miR160 mature sequence is highly conserved in various species, and the miR160 mature sequence formed by the 5' arm of the MIR160 precursor was more conserved. Three FveARFs in woodland strawberry were negatively regulated by FvemiR160a, among which FveARF18A was the most significant. Phylogenetic analysis indicated that FvemiR160 is closely related to apple (Malus domestica), grape (Vitis vinifera), and Arabidopsis thaliana, while FveARF18A is closely related to RcARF18. Subsequently, we demonstrated that FvemiR160a can target cutting FveARF18A to negatively regulate its expression by RLM-5' RACE, cleavage site mutation, and GFP fluorescence assay. Moreover, we observed that FveMIR160a overexpressed plants have advanced flowering, while mFveARF18A overexpressed plants have delayed flowering. We also verified that FveARF18A negatively regulates the expression of FveAP1 and FveFUL by binding their promoters by yeast one-hybrid, LUC, and GUS assay, and FveAP1 and FveFUL transgenic Arabidopsis showed early flowering phenotype. In addition, the expression level of FvemiR160a was decreased obviously while that of FveARF18A was increased obviously by MeJA, GA and IAA. In conclusion, our study reveals the important role of the FvemiR160-FveARF18A-FveAP1/FveFUL module in the flowering process of woodland strawberry and provides a new pathway for studying flowering.


Subject(s)
Fragaria , Fragaria/genetics , Fragaria/metabolism , Phylogeny , Plant Leaves/genetics , Phenotype , Promoter Regions, Genetic , Gene Expression Regulation, Plant/genetics
5.
J Virol ; : e0052124, 2024 Jun 14.
Article in English | MEDLINE | ID: mdl-38874361

ABSTRACT

The reoccurrence of successive waves of SARS-CoV-2 variants suggests the exploration of more vaccine alternatives is imperative. Modified vaccinia virus Ankara (MVA) is a virus vector exhibiting excellent safety as well as efficacy for vaccine development. Here, a series of recombinant MVAs (rMVAs) expressing monomerized or trimerized S proteins from different SARS-CoV-2 variants are engineered. Trimerized S expressed from rMVAs is found predominantly as trimers on the surface of infected cells. Remarkably, immunization of mice with rMVAs demonstrates that S expressed in trimer elicits higher levels of binding IgG and IgA, as well as neutralizing antibodies for matched and mismatched S proteins than S in the monomer. In addition, trimerized S expressed by rMVA induces enhanced cytotoxic T-cell responses than S in the monomer. Importantly, the rMVA vaccines expressing trimerized S exhibit superior protection against a lethal SARS-CoV-2 challenge as the immunized animals all survive without displaying any pathological conditions. This study suggests that opting for trimerized S may represent a more effective approach and highlights that the MVA platform serves as an ideal foundation to continuously advance SARS-CoV-2 vaccine development. IMPORTANCE: MVA is a promising vaccine vector and has been approved as a vaccine for smallpox and mpox. Our analyses suggested that recombinant MVA expressing S in trimer (rMVA-ST) elicited robust cellular and humoral immunity and was more effective than MVA-S-monomer. Importantly, the rMVA-ST vaccine was able to stimulate decent cross-reactive neutralization against pseudoviruses packaged using S from different sublineages, including Wuhan, Delta, and Omicron. Remarkably, mice immunized with rMVA-ST were completely protected from a lethal challenge of SARS-CoV-2 without displaying any pathological conditions. Our results demonstrated that an MVA vectored vaccine expressing trimerized S is a promising vaccine candidate for SARS-CoV-2 and the strategy might be adapted for future vaccine development for coronaviruses.

6.
Mol Ecol ; : e17356, 2024 Apr 18.
Article in English | MEDLINE | ID: mdl-38634782

ABSTRACT

DNA methylation has been proposed to be an important mechanism that allows plants to respond to their environments sometimes entirely uncoupled from genetic variation. To understand the genetic basis, biological functions and climatic relationships of DNA methylation at a population scale in Arabidopsis thaliana, we performed a genome-wide association analysis with high-quality single nucleotide polymorphisms (SNPs), and found that ~56% on average, especially in the CHH sequence context (71%), of the differentially methylated regions (DMRs) are not tagged by SNPs. Among them, a total of 3235 DMRs are significantly associated with gene expressions and potentially heritable. 655 of the 3235 DMRs are associated with climatic variables, and we experimentally verified one of them, HEI10 (HUMAN ENHANCER OF CELL INVASION NO.10). Such epigenetic loci could be subjected to natural selection thereby affecting plant adaptation, and would be expected to be an indicator of accessions at risk. We therefore incorporated these climate-related DMRs into a gradient forest model, and found that the natural A. thaliana accessions in Southern Europe that may be most at risk under future climate change. Our findings highlight the importance of integrating DNA methylation that is independent of genetic variations, and climatic data to predict plants' vulnerability to future climate change.

7.
Plant Physiol ; 191(2): 1344-1364, 2023 02 12.
Article in English | MEDLINE | ID: mdl-36417260

ABSTRACT

Wheat (Triticum aestivum) is particularly susceptible to water deficit at the jointing stage of its development. Sucrose non-fermenting 1-related protein kinase 2 (SnRK2) acts as a signaling hub in the response to drought stress, but whether SnRK2 helps plants cope with water deficit via other mechanisms is largely unknown. Here, we cloned and characterized TaSnRK2.10, which was induced by multiple abiotic stresses and phytohormones. Ectopic expression of TaSnRK2.10 in rice (Oryza sativa) conferred drought tolerance, manifested by multiple improved physiological indices, including increased water content, cell membrane stability, and survival rates, as well as decreased water loss and accumulation of H2O2 and malonaldehyde. TaSnRK2.10 interacted with and phosphorylated early responsive to dehydration 15 (TaERD15) and enolase 1 (TaENO1) in vivo and in vitro. TaERD15 phosphorylated by TaSnRK2.10 was prone to degradation by the 26S proteasome, thereby mitigating its negative effects on drought tolerance. Phosphorylation of TaENO1 by TaSnRK2.10 may account for the substantially increased levels of phosphoenolpyruvate (PEP), a key metabolite of primary and secondary metabolism, in TaSnRK2.10-overexpressing rice, thereby enhancing its viability under drought stress. Our results demonstrate that TaSnRK2.10 not only regulated stomatal aperture and the expression of drought-responsive genes, but also enhanced PEP supply and promoted the degradation of TaERD15, all of which enhanced drought tolerance.


Subject(s)
Oryza , Triticum , Triticum/metabolism , Oryza/genetics , Oryza/metabolism , Drought Resistance , Plant Proteins/metabolism , Hydrogen Peroxide/metabolism , Droughts , Water/metabolism , Stress, Physiological/genetics , Plants, Genetically Modified/metabolism , Gene Expression Regulation, Plant
8.
Opt Express ; 32(9): 14953-14962, 2024 Apr 22.
Article in English | MEDLINE | ID: mdl-38859158

ABSTRACT

In this work, we hybridize an air cavity reflector and a nanopatterned sapphire substrate (NPSS) for making an inclined-sidewall-shaped deep ultraviolet micro light-emitting diode (DUV micro-LED) array to enhance the light extraction efficiency (LEE). A cost-effective hybrid photolithography process involving positive and negative photoresist (PR) is explored to fabricate air-cavity reflectors. The experimental results demonstrate a 9.88% increase in the optical power for the DUV micro-LED array with a bottom air-cavity reflector when compared with the conventional DUV micro-LED array with only a sidewall metal reflector. The bottom air-cavity reflector significantly contributes to the reduction of the light absorption and provides more escape paths for light, which in turn increases the LEE. Our investigations also report that such a designed air-cavity reflector exhibits a more pronounced impact on small-size micro-LED arrays, because more photons can propagate into escape cones by experiencing fewer scattering events from the air-cavity structure. Furthermore, the NPSS can enlarge the escape cone and serve as scattering centers to eliminate the waveguiding effect, which further enables the improved LEE for the DUV micro-LED array with an air-cavity reflector.

9.
Opt Lett ; 49(11): 3275-3278, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38824382

ABSTRACT

It is known that light extraction efficiency (LEE) for AlGaN-based deep ultraviolet (DUV) light-emitting diodes (LEDs) can be enhanced by using an inclined sidewall of mesa. However, the reported optimal inclined angles are different. In this work, to explore the origin for enhancing the LEE of DUV LED by using inclined sidewalls, we investigate the effect of an inclined sidewall angle on the LEE for AlGaN-based DUV LEDs with different mesa diameters by using ray tracing. It is found that when compared to large-size DUV LEDs with inclined sidewall, the LEE of small-size DUV LEDs with inclined sidewall is enhanced from both the bottom and side surfaces due to the reduced scattering length and material absorption. Additionally, the optimal inclined sidewall angle is recommended within the range of 25°-65°, and the optimal angle for DUV LEDs decreases as the chip size increases. It can be attributed to the fact that there are two scattering mechanisms for the inclined sidewall. For smaller chip sizes, most of the light is directly scattered into escape cones by the inclined sidewall, resulting in a larger optimal angle. For larger chip sizes, the light firstly experiences total internal reflections by the out-light plane and then is scattered into escape cones by the inclined sidewalls, leading to a smaller optimal angle.

10.
Opt Lett ; 49(11): 3279-3282, 2024 Jun 01.
Article in English | MEDLINE | ID: mdl-38824383

ABSTRACT

AlGaN-based solar-blind ultraviolet avalanche detectors have huge potentials in the fields of corona discharge monitoring, biological imaging, etc. Here, we study the impact of the heterojunction polarization-related effects on the AlGaN-based solar-blind ultraviolet avalanche detectors. Our work confirms that the polarization heterojunction is beneficial to reducing avalanche bias and lifting avalanche gain by improving the electric field in the depletion region, while the polarization-induced fixed charges will lead to a redistribution of the electrons, in turn shielding the charges and weakening the electric field enhancement effect. This shielding effect will need external bias to eliminate, and that is why the polarization heterojunction cannot work at relatively low bias but has an enhancement effect at high bias. Controlling the doping level between the hetero-interface can affect the shielding effect. An unintentionally doped polarization heterojunction can effectively reduce the shielding effect, thus reducing the avalanche bias. The conclusions also hold true for the negative polarization regime. We believe our findings can provide some useful insights for the design of the AlGaN-based solar-blind ultraviolet detectors.

11.
J Transl Med ; 21(1): 347, 2023 05 25.
Article in English | MEDLINE | ID: mdl-37231493

ABSTRACT

Cardiovascular disease (CVD) is the leading cause of noncommunicable disease-related death worldwide, and effective therapeutic strategies against CVD are urgently needed. Mitochondria dysfunction involves in the onset and development of CVD. Nowadays, mitochondrial transplantation, an alternative treatment aimed at increasing mitochondrial number and improving mitochondrial function, has been emerged with great therapeutic potential. Substantial evidence indicates that mitochondrial transplantation improves cardiac function and outcomes in patients with CVD. Therefore, mitochondrial transplantation has profound implications in the prevention and treatment of CVD. Here, we review the mitochondrial abnormalities that occur in CVD and summarize the therapeutic strategies of mitochondrial transplantation for CVD.


Subject(s)
Cardiovascular Diseases , Humans , Cardiovascular Diseases/therapy , Mitochondria
12.
Opt Express ; 31(2): 2931-2941, 2023 Jan 16.
Article in English | MEDLINE | ID: mdl-36785295

ABSTRACT

In this work, by using three-dimensional finite-difference time-domain (3D FDTD) method, the effect of conventional nano-patterned sapphire substrate (NPSS) on the optical crosstalk and the light extraction efficiency (LEE) for InGaN/GaN-based flip-chip micro light-emitting diodes (µ-LEDs) are systematically studied. We find that the conventional NPSS is not suitable for µ-LEDs. It is because the inclined mesa sidewall for µ-LEDs possesses a good scattering effect for µ-LEDs, but the introduced conventional NPSS causes part of the light be off escape cone between sapphire and air and become the guided light. To suppress the guided light and improve the optical crosstalk, a thick air layer between the n-GaN layer and the sapphire substrate can be used as a light filter to prevent the guided light from propagating into the sapphire. However, in reality, it is challenging to make the aforementioned air layer from point of fabrication view. Therefore, we propose the air-cavity patterned sapphire substrate (AC-PSS) as the light filter. Our results show that the crosstalk ratio can be decreased to the value even lower than 10%. The LEE can also be enhanced simultaneously due to combination effects of the filtering effect of the AC-PSS and the scattering effect of the inclined mesa sidewall.

13.
Opt Lett ; 48(22): 5863-5866, 2023 Nov 15.
Article in English | MEDLINE | ID: mdl-37966738

ABSTRACT

In this Letter, beveled mesas for 30 × 30 µm2 GaN-based micro-light-emitting diodes (µLEDs) with different inclination angles are designed, fabricated, and measured. We find that µLED with a mesa inclination angle of 28° has the lowest internal quantum efficiency (IQE) and the highest injection current density at which the peak IQE is obtained. This is due to the increased quantum confined Stark effect (QCSE) at the mesa edge. The increased QCSE results from the strong electric field coupling effect. Instead of radiative recombination, more nonradiative recombination and leakage current will be generated in the sidewall regions. Besides, the smallest angle (28°) also produces the lowest light extraction efficiency (LEE), which arises from the optical loss caused by the sidewall reflection at the beveled surface sides. Therefore, the inclination angle for the beveled mesa has to be increased to 52° and 61° by using Ni and SiO2 as hard masks, respectively. Experimental and numerical results show that the external quantum efficiency (EQE) and the optical power can be enhanced for the fabricated devices. Meanwhile, the reduced surface recombination rate also decreases the leakage current.

14.
Pulm Pharmacol Ther ; 80: 102202, 2023 06.
Article in English | MEDLINE | ID: mdl-36906117

ABSTRACT

Pulmonary fibrosis is a chronic interstitial fibrosis lung disease with high mortality, which is often complicated with lung cancer. The incidence of IPF complicated with lung cancer is getting higher and higher. At present, there is no consensus on the management and treatment of pulmonary fibrosis patients with lung cancer. There is an urgent need to develop preclinical drug evaluation methods for IPF with lung cancer and potential therapeutic drugs for IPF with lung cancer. The pathogenic mechanism of IPF is similar to that of lung cancer, and the multi-effect drugs with anticancer and anti-fibrosis will have potential value in the treatment of IPF complicated with lung cancer. In this study, we established an animal model of IPF complicated with lung cancer in situ to evaluate the therapeutic effect of the antiangiogenic drug anlotinib. The pharmacodynamic results in vivo showed that anlotinib could significantly improve the lung function of IPF-LC mice, reduce the content of collagen in lung tissue, increase the survival rate of mice, and inhibit the growth of lung tumor in mice. The results of Western blot and immunohistochemical analysis of lung tissue showed that anlotinib significantly inhibited the expression of fibrosis marker protein α-SMA, Collagen I and Fibronectin and tumor proliferation marker protein PCNA in mouse lung tissue, and down-regulated the content of serum tumor marker CEA. Through transcriptome analysis, we found that anlotinib regulates MAPK signal pathway, PARP signal pathway and coagulation cascade signal pathway in lung cancer and pulmonary fibrosis, which all play an important role in lung cancer and pulmonary fibrosis. In addition, there is crosstalk between the signal pathway participated by the target of anlotinib and MAPK, JAK/STAT and mTOR signal pathway. In summary, anlotinib will be a candidate for IPF-LC treatment.


Subject(s)
Adenocarcinoma of Lung , Idiopathic Pulmonary Fibrosis , Lung Diseases, Interstitial , Lung Neoplasms , Mice , Animals , Idiopathic Pulmonary Fibrosis/complications , Idiopathic Pulmonary Fibrosis/drug therapy , Idiopathic Pulmonary Fibrosis/metabolism , Lung , Lung Neoplasms/complications , Lung Neoplasms/drug therapy , Lung Neoplasms/metabolism , Lung Diseases, Interstitial/pathology , Adenocarcinoma of Lung/drug therapy , Collagen/metabolism , Biomarkers/metabolism , Bleomycin/pharmacology
15.
Analyst ; 148(24): 6341-6349, 2023 Dec 04.
Article in English | MEDLINE | ID: mdl-37955601

ABSTRACT

SO2, NO2 and NO are the main atmospheric pollutants produced by the combustion of fossil fuel. Detecting these gases is of great significance for atmospheric protection and the online concentration detection of pollutants. In this study, the concentration retrieval methods of NO, NO2 and SO2 and their mutual effects were studied in the wavelength range of 192.3-254.4 nm. In this band, NO, NO2 and SO2 have large absorption cross-sections; however, their spectrum superpositions were serious. A novel method was proposed to separate the superposed absorption spectra of NO and SO2 or NO2. The advantage of this method is that it can remove the influence of SO2 and NO2 on NO concentration retrieval. The fast Fourier transform (FFT) amplitude method was used to calculate the concentrations of SO2 and NO2, and the direct absorption spectroscopy method was used to calculate NO concentration. Via these methods, the gas concentrations of SO2, NO2 and NO can be calculated in ternary-gas mixtures. The experimental results show that these methods can effectively remove the mutual interferences between the concentration retrieval of NO, NO2 and SO2. The maximum absolute values of the relative deviations for the concentration retrieval of SO2, NO2 and NO in ternary-gas mixtures are 3.868%, 4.740% and 5.008%, respectively. These methods have high detection precision and good adaptability and are suitable for online flue detection equipment.

16.
Appl Opt ; 62(13): 3431-3438, 2023 May 01.
Article in English | MEDLINE | ID: mdl-37132844

ABSTRACT

In this paper, by using advanced numerical models, we investigate the impact of the AlN/GaN distributed Bragg reflector (DBR) and AlInN/GaN DBR on stimulated radiative recombination for GaN-based vertical-cavity-surface-emitting lasers (VCSELs). According to our results, when compared with the VCSEL with AlN/GaN DBR, we find that the VCSEL with AlInN/GaN DBR decreases the polarization-induced electric field in the active region, and this helps to increase the electron-hole radiative recombination. However, we also find that the AlInN/GaN DBR has a reduced reflectivity when compared with the AlN/GaN DBR with the same number of pairs. Furthermore, this paper suggests that more pairs of AlInN/GaN DBR will be set, which helps to even further increase the laser power. Hence, the 3 dB frequency can be increased for the proposed device. In spite of the increased laser power, the smaller thermal conductivity for AlInN than AlN results in the earlier thermal droop in the laser power for the proposed VCSEL.

17.
Biotechnol Lett ; 45(1): 115-124, 2023 Jan.
Article in English | MEDLINE | ID: mdl-36450976

ABSTRACT

OBJECTIVE: To examine the influence of widely used protein affinity tags and the tobacco PR1a signal peptide (SP) on detection, purification and bioactivity analyses of the small oomycete apoplastic effector SCR96 in planta. RESULTS: Through agroinfiltration, the phytotoxic effector SCR96 of Phytophthora cactorum was expressed in Nicotiana benthamiana leaf apoplast as a fusion protein carrying single affinity tag (His, HA or FLAG) at either C- or N-terminus. Leaf necrosis caused by different affinity-tagged SCR96 varied among tags and replicates. All of tagged proteins can be detected by antibodies against SCR96. All of SCR96 fusions except N-terminally fused 6His-tagged protein were detected using tag antibodies, indicating that 6His tag may be degraded when fused at N-terminus. Interestingly, C-terminal His- and FLAG-tagged SCR96 maintained the biological activity after purification. In the substitution assay of SCR96 SP, we observed that PR1a SP can lead chimeric SCR96 expression in N. benthamiana, but the replacement totally disrupted its bioactivity. CONCLUSION: C-terminal His or FLAG tag, along with its original SP, is efficient enough to enable detection and purification of functional SCR96 from N. benthamiana leaf apoplast, which would facilitate plant-pathogen interaction studies.


Subject(s)
Nicotiana , Phytophthora , Nicotiana/genetics , Nicotiana/metabolism , Protein Sorting Signals/genetics , Proteins/metabolism , Phytophthora/genetics , Phytophthora/metabolism , Antibodies/metabolism , Chromatography, Affinity
18.
Sensors (Basel) ; 23(10)2023 May 10.
Article in English | MEDLINE | ID: mdl-37430522

ABSTRACT

Spectral reflectance or transmittance measurements provide intrinsic information on the material of an object and are widely used in remote sensing, agriculture, diagnostic medicine, etc. Most reconstruction-based spectral reflectance or transmittance measurement methods based on broadband active illumination use narrow-band LEDs or lamps combined with specific filters as spectral encoding light sources. These light sources cannot achieve the designed spectral encoding with a high resolution and accuracy due to their low degree of freedom for adjustment, leading to inaccurate spectral measurements. To address this issue, we designed a spectral encoding simulator for active illumination. The simulator is composed of a prismatic spectral imaging system and a digital micromirror device. The spectral wavelengths and intensity are adjusted by switching the micromirrors. We used it to simulate spectral encodings according to the spectral distribution on micromirrors and solved the DMD patterns corresponding to the spectral encodings with a convex optimization algorithm. To verify the applicability of the simulator for spectral measurements based on active illumination, we used it to numerically simulate existing spectral encodings. We also numerically simulated a high-resolution Gaussian random measurement encoding for compressed sensing and measured the spectral reflectance of one vegetation type and two minerals through numerical simulations. We reconstructed the spectral transmittance of a calibrated filter through an experiment. The results show that the simulator can measure the spectral reflectance or transmittance with a high resolution and accuracy.

19.
J Biol Chem ; 296: 100130, 2021.
Article in English | MEDLINE | ID: mdl-33262216

ABSTRACT

Meiosis, which produces haploid progeny, is critical to ensuring both faithful genome transmission and genetic diversity. Proteasomes play critical roles at various stages of spermatogenesis, including meiosis, but the underlying mechanisms remain unclear. The atypical proteasomes, which contain the activator PA200, catalyze the acetylation-dependent degradation of the core histones in elongated spermatids and DNA repair in somatic cells. We show here that the testis-specific proteasome subunit α4s/PSMA8 is essential for male fertility by promoting proper formation of spermatoproteasomes, which harbor both PA200 and constitutive catalytic subunits. Immunostaining of a spermatocyte marker, SYCP3, indicated that meiosis was halted at the stage of spermatocytes in the α4s-deficient testes. α4s stimulated the in vitro degradation of the acetylated core histones, instead of nonacetylated histones, by the PA200-proteasome. Deletion of α4s blocked degradation of the core histones at DNA damage loci in spermatocytes, leading to meiotic arrest at metaphase I. Thus, α4s is required for histone degradation at meiotic DNA damage loci, proper progression of meiosis, and fertility in males by promoting proper formation of spermatoproteasomes. These results are important for understanding male infertility and might provide potential targets for male contraception or treatment of male infertility.


Subject(s)
DNA Repair , Histones/metabolism , Infertility, Male/pathology , Meiosis , Proteasome Endopeptidase Complex/metabolism , Spermatocytes/cytology , Spermatogenesis , Animals , DNA Damage , Infertility, Male/etiology , Infertility, Male/metabolism , Male , Mice , Mice, Inbred C57BL , Proteasome Endopeptidase Complex/genetics , Spermatids , Spermatocytes/metabolism
20.
Plant Biotechnol J ; 20(6): 1122-1139, 2022 06.
Article in English | MEDLINE | ID: mdl-35189026

ABSTRACT

Melatonin, a natural phytohormone in plants, plays multiple critical roles in plant growth and stress responses. Although melatonin biosynthesis-related genes have been suggested to possess diverse biological functions, their roles and functional mechanisms in regulating rice grain yield remain largely unexplored. Here, we uncovered the roles of a caffeic acid O-methyltransferase (OsCOMT) gene in mediating rice grain yield through dual regulation of leaf senescence and vascular development. In vitro and in vivo evidence revealed that OsCOMT is involved in melatonin biosynthesis. Transgenic assays suggested that OsCOMT significantly delays leaf senescence at the grain filling stage by inhibiting degradation of chlorophyll and chloroplast, which, in turn, improves photosynthesis efficiency. In addition, the number and size of vascular bundles in the culms and leaves were significantly increased in the OsCOMT-overexpressing plants, while decreased in the knockout plants, suggesting that OsCOMT plays a positive role in vascular development of rice. Further evidence indicated that OsCOMT-mediated vascular development might owe to the crosstalk between melatonin and cytokinin. More importantly, we found that OsCOMT is a positive regulator of grain yield, and overexpression of OsCOMT increase grain yield per plant even in a high-yield variety background, suggesting that OsCOMT can be used as an important target for enhancing rice yield. Our findings shed novel insights into melatonin-mediated leaf senescence and vascular development and provide a possible strategy for genetic improvement of rice grain yield.


Subject(s)
Melatonin , Oryza , Edible Grain , Gene Expression Regulation, Plant/genetics , Melatonin/genetics , Melatonin/metabolism , Methyltransferases , Oryza/metabolism , Plant Leaves/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Senescence
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