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1.
Plant Cell ; 34(11): 4366-4387, 2022 10 27.
Article in English | MEDLINE | ID: mdl-35972379

ABSTRACT

Ethylene plays essential roles in adaptive growth of rice (Oryza sativa). Understanding of the crosstalk between ethylene and auxin (Aux) is limited in rice. Here, from an analysis of the root-specific ethylene-insensitive rice mutant mao hu zi 10 (mhz10), we identified the tryptophan aminotransferase (TAR) MHZ10/OsTAR2, which catalyzes the key step in indole-3-pyruvic acid-dependent Aux biosynthesis. Genetically, OsTAR2 acts downstream of ethylene signaling in root ethylene responses. ETHYLENE INSENSITIVE3 like1 (OsEIL1) directly activated OsTAR2 expression. Surprisingly, ethylene induction of OsTAR2 expression still required the Aux pathway. We also show that Os indole-3-acetic acid (IAA)1/9 and OsIAA21/31 physically interact with OsEIL1 and show promotive and repressive effects on OsEIL1-activated OsTAR2 promoter activity, respectively. These effects likely depend on their EAR motif-mediated histone acetylation/deacetylation modification. The special promoting activity of OsIAA1/9 on OsEIL1 may require both the EAR motifs and the flanking sequences for recruitment of histone acetyltransferase. The repressors OsIAA21/31 exhibit earlier degradation upon ethylene treatment than the activators OsIAA1/9 in a TIR1/AFB-dependent manner, allowing OsEIL1 activation by activators OsIAA1/9 for OsTAR2 expression and signal amplification. This study reveals a positive feedback regulation of ethylene signaling by Aux biosynthesis and highlights the crosstalk between ethylene and Aux pathways at a previously underappreciated level for root growth regulation in rice.


Subject(s)
Ethylenes , Indoleacetic Acids , Oryza , Plant Roots , Tryptophan Transaminase , Ethylenes/metabolism , Gene Expression Regulation, Plant , Indoleacetic Acids/metabolism , Oryza/growth & development , Oryza/metabolism , Plant Roots/growth & development , Plant Roots/metabolism , Tryptophan Transaminase/genetics , Tryptophan Transaminase/metabolism
2.
Mol Ther ; 32(8): 2461-2469, 2024 Aug 07.
Article in English | MEDLINE | ID: mdl-38796701

ABSTRACT

N6-methyladenosine (m6A) is the most abundant endogenous modification in eukaryotic RNAs. It plays important roles in various biological processes and diseases, including cancers. More and more studies have revealed that the deposition of m6A is specifically regulated in a context-dependent manner. Here, we review the diverse mechanisms that determine the topology of m6A along RNAs and the cell-type-specific m6A methylomes. The exon junction complex (EJC) as well as histone modifications play important roles in determining the topological distribution of m6A along nascent RNAs, while the transcription factors and RNA-binding proteins, which usually bind specific DNAs and RNAs in a cell-type-specific manner, largely account for the cell-type-specific m6A methylomes. Due to the lack of specificity of m6A writers and readers, there are still challenges to target the core m6A machinery for cancer therapies. Therefore, understanding the mechanisms underlying the specificity of m6A modifications in cancers would be important for future cancer therapies through m6A intervention.


Subject(s)
Adenosine , Neoplasms , Humans , Neoplasms/genetics , Neoplasms/therapy , Neoplasms/metabolism , Adenosine/analogs & derivatives , Adenosine/metabolism , Methylation , RNA/metabolism , RNA/genetics , Animals , RNA-Binding Proteins/metabolism , RNA-Binding Proteins/genetics , Gene Expression Regulation, Neoplastic , RNA Methylation
3.
J Biol Chem ; 299(6): 104780, 2023 06.
Article in English | MEDLINE | ID: mdl-37142220

ABSTRACT

The calcium-activated chloride channel TMEM16A is a potential drug target to treat hypertension, secretory diarrhea, and several cancers. However, all reported TMEM16A structures are either closed or desensitized, and direct inhibition of the open state by drug molecules lacks a reliable structural basis. Therefore, revealing the druggable pocket of TMEM16A exposed in the open state is important for understanding protein-ligand interactions and facilitating rational drug design. Here, we reconstructed the calcium-activated open conformation of TMEM16A using an enhanced sampling algorithm and segmental modeling. Furthermore, we identified an open-state druggable pocket and screened a potent TMEM16A inhibitor, etoposide, which is a derivative of a traditional herbal monomer. Molecular simulations and site-directed mutagenesis showed that etoposide binds to the open state of TMEM16A, thereby blocking the ion conductance pore of the channel. Finally, we demonstrated that etoposide can target TMEM16A to inhibit the proliferation of prostate cancer PC-3Ā cells. Together, these findings provide a deep understanding of the TMEM16A open state at an atomic level and identify pockets for the design of novel inhibitors with broad applications in chloride channel biology, biophysics, and medicinal chemistry.


Subject(s)
Anoctamin-1 , Models, Molecular , Humans , Male , Anoctamin-1/chemistry , Anoctamin-1/metabolism , Calcium/metabolism , Etoposide/pharmacology , Protein Binding , Protein Structure, Tertiary , Cell Line, Tumor , Cell Proliferation/drug effects , Computer Simulation
4.
New Phytol ; 243(5): 1724-1741, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38509454

ABSTRACT

Gene expression is regulated at multiple levels, including RNA processing and DNA methylation/demethylation. How these regulations are controlled remains unclear. Here, through analysis of a suppressor for the OsEIN2 over-expressor, we identified an RNA recognition motif protein SUPPRESSOR OF EIN2 (SOE). SOE is localized in nuclear speckles and interacts with several components of the spliceosome. We find SOE associates with hundreds of targets and directly binds to a DNA glycosylase gene DNG701 pre-mRNA for efficient splicing and stabilization, allowing for subsequent DNG701-mediated DNA demethylation of the transgene promoter for proper gene expression. The V81M substitution in the suppressor mutant protein mSOE impaired its protein stability and binding activity to DNG701 pre-mRNA, leading to transgene silencing. SOE mutation enhances grain size and yield. Haplotype analysis in c. 3000 rice accessions reveals that the haplotype 1 (Hap 1) promoter is associated with high 1000-grain weight, and most of the japonica accessions, but not indica ones, have the Hap 1 elite allele. Our study discovers a novel mechanism for the regulation of gene expression and provides an elite allele for the promotion of yield potentials in rice.


Subject(s)
Gene Expression Regulation, Plant , Gene Silencing , Oryza , Plant Proteins , Transgenes , Oryza/genetics , Plant Proteins/genetics , Plant Proteins/metabolism , Promoter Regions, Genetic/genetics , Mutation/genetics , Protein Domains , Haplotypes/genetics , DNA Methylation/genetics , Protein Binding , Plants, Genetically Modified , Alleles
5.
Arch Virol ; 169(6): 119, 2024 May 16.
Article in English | MEDLINE | ID: mdl-38753197

ABSTRACT

Porcine circovirus (PCV) has become a major pathogen, causing major economic losses in the global pig industry, and PCV type 2 (PCV2) and 3 (PCV3) are distributed worldwide.Ā We designed specific primer and probe sequences targeting PCV2 Cap and PCV3 Rap andĀ developed a multiplex crystal digital PCR (cdPCR) method after optimizing the primerĀ concentration, probe concentration, and annealing temperature.Ā The multiplex cdPCR assay permits precise and differential detection of PCV2 and PCV3, with a limit of detection of 1.39 Ɨ 101 and 1.27 Ɨ 101 copies/reaction, respectively, and no cross-reaction with other porcine viruses was observed. The intra-assay and interassay coefficients of variation (CVs) were less than 8.75%, indicating good repeatability and reproducibility. To evaluate the practical value of this assay, 40 tissue samples and 70 feed samples were tested for both PCV2 and PCV3 by cdPCR and quantitative PCR (qPCR). Using multiplex cdPCR, the rates of PCV2 infection, PCV3 infection, and coinfection were 28.45%, 1.72%, and 12.93%, respectively, and using multiplex qPCR, they were 25.00%, 0.86%, and 4.31%, respectively This highly specific and sensitive multiplex cdPCR thus allows accurate simultaneous detection of PCV2 and PCV3, and it is particularly well suited for applications that require the detection of small amounts of input nucleic acid or samples with intensive processing and complex matrices.


Subject(s)
Circoviridae Infections , Circovirus , Multiplex Polymerase Chain Reaction , Swine Diseases , Circovirus/genetics , Circovirus/isolation & purification , Circovirus/classification , Swine , Animals , Circoviridae Infections/veterinary , Circoviridae Infections/virology , Circoviridae Infections/diagnosis , Swine Diseases/virology , Swine Diseases/diagnosis , Multiplex Polymerase Chain Reaction/methods , Sensitivity and Specificity , Reproducibility of Results , DNA Primers/genetics , DNA, Viral/genetics
6.
Mol Ther ; 31(7): 2089-2104, 2023 07 05.
Article in English | MEDLINE | ID: mdl-36945773

ABSTRACT

CAR TĀ cells recognizing CD19 effectively treat relapsed and refractory B-ALL and DLBCL. However, CD19 loss is a frequent cause of relapse. Simultaneously targeting a second antigen, CD22, may decrease antigen escape, but is challenging: its density is approximately 10-fold less than CD19, and its large structure may hamper immune synapse formation. The characteristics of the optimal CD22 CAR are underexplored. We generated 12 distinct CD22 antibodies and tested CARs derived from them to identify a CAR based on the novel 9A8 antibody, which was sensitive to low CD22 density and lacked tonic signaling. We found no correlation between affinity or membrane proximity of recognition epitope within Ig domains 3-6 of CD22 with CART function. The optimal strategy for CD19/CD22 CART co-targeting is undetermined. Co-administration of CD19 and CD22 CARs is costly; single CARs targeting CD19 and CD22 are challenging to construct. The co-expression of two CARs has previously been achieved using bicistronic vectors. Here, we generated a dual CART product by co-transduction with 9A8-41BBƎĀ¶ and CAT-41BBƎĀ¶ (obe-cel), the previously described CD19 CAR. CAT/9A8 CART eliminated single- and double-positive target cells inĀ vitro and eliminated CD19- tumors inĀ vivo. CAT/9A8 CART is being tested in a phase I clinical study (NCT02443831).


Subject(s)
Burkitt Lymphoma , Receptors, Chimeric Antigen , Humans , Receptors, Chimeric Antigen/metabolism , T-Lymphocytes , Neoplasm Recurrence, Local , Immunotherapy, Adoptive , Adaptor Proteins, Signal Transducing , Antigens, CD19 , Antibodies , Sialic Acid Binding Ig-like Lectin 2
7.
Phytother Res ; 2024 Aug 09.
Article in English | MEDLINE | ID: mdl-39120263

ABSTRACT

Women go through several predictable conditions and symptoms during menopause that are caused by age, changes in sex hormone levels, and other factors. Conventional menopause hormone therapy has raised serious concerns about the increased risks of cancers, blood clots, depression, etc. Selective estrogen receptor modulators (SERMs) that can be both agonists and antagonists of estrogen receptors in a tissue-specific manner are being developed to reduce the health concerns associated with menopause hormone therapy. Here, we have searched the Chinese national traditional Chinese medicine (TCM) patent database to identify potential SERM-like compounds with reduced health risks. TCM has been widely used for treating complex symptoms associated with menopause syndrome and thus can be a particularly rich source for pharmaceutical alternatives with SERM properties. After extensive literature review and molecular simulation, we conclude that protopanaxatriol, paeoniflorin, astragalin, catalpol, and hyperoside among others may be particularly promising as SERM-like compounds in treating the menopausal syndrome. Compounds in TCM hold promise in yielding comparable outcomes to hormone therapy but with reduced associated risks, thus presenting promising avenues for their clinical applications.

8.
Phytochem Anal ; 35(5): 1174-1185, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38553821

ABSTRACT

INTRODUCTION: Sainfoin (Onobrychis viciaefolia) is a vital legume forage, and drought is the primary element impeding sainfoin growth. OBJECTIVE: The anatomical structure, physiological indexes, and metabolites of the leaves of sainfoin seedlings with a drought-resistant line of P1 (DRL) and a drought-sensitive material of 2049 (DSM) were analyzed under drought (-1.0 MPa) with polyethylene glycol-6000 (PEG-6000). METHODS: The leaf anatomy was studied by the paraffin section method. The related physiological indexes were measured by the hydroxylamine oxidation method, titanium sulfate colorimetric method, thiobarbituric acid method, acidic ninhydrin colorimetric method, and Coomassie brilliant blue method. The metabolomics analysis was composed of liquid chromatography tandem high-resolution mass spectrometry (LC-MS/MS). RESULTS: The results revealed that the thickness of the epidermis, palisade tissue, and sponge tissue of DRL were significantly greater than those of DSM. The leaves of DRL exhibited lower levels of superoxide anion (O2 Ć¢Ā€Ā¢-) production rate, hydrogen peroxide (H2O2) content, and malondialdehyde (MDA) content compared with DSM, while proline (Pro) content and soluble protein (SP) content were significantly higher than those of DSM. A total of 391 differential metabolites were identified in two samples. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment showed that the primary differential metabolites were concentrated into the tyrosine metabolism; isoquinoline alkaloid biosynthesis; ubiquinone and other terpenoid quinone biosynthesis; neomycin, kanamycin, and gentamicin biosynthesis; and anthocyanin biosynthesis metabolic pathways. CONCLUSION: Compared with DSM, DRL had more complete anatomical structure, lower active oxygen content, and higher antioxidant level. The results improved our insights into the drought-resistant mechanisms in sainfoin.


Subject(s)
Droughts , Metabolome , Plant Leaves , Seedlings , Plant Leaves/metabolism , Plant Leaves/anatomy & histology , Seedlings/metabolism , Fabaceae/physiology , Fabaceae/anatomy & histology , Fabaceae/metabolism , Stress, Physiological , Metabolomics
9.
Int J Mol Sci ; 25(18)2024 Sep 23.
Article in English | MEDLINE | ID: mdl-39337671

ABSTRACT

Neurodegenerative diseases are the leading cause of human disability and immensely reduce patients' life span and quality. The diseases are characterized by the functional loss of neuronal cells and share several common pathogenic mechanisms involving the malfunction, structural distortion, or aggregation of multiple key regulatory proteins. Cellular phase separation is the formation of biomolecular condensates that regulate numerous biological processes, including neuronal development and synaptic signaling transduction. Aberrant phase separation may cause protein aggregation that is a general phenomenon in the neuronal cells of patients suffering neurodegenerative diseases. In this review, we summarize the pathological causes of common neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, and Huntington's disease, among others. We discuss the regulation of key amyloidogenic proteins with an emphasis of their aberrant phase separation and aggregation. We also introduce the approaches as potential therapeutic strategies to ameliorate neurodegenerative diseases through intervening protein aggregation. Overall, this review consolidates the research findings of phase separation and aggregation caused by misfolded proteins in a context of neurodegenerative diseases.


Subject(s)
Neurodegenerative Diseases , Protein Aggregation, Pathological , Humans , Neurodegenerative Diseases/metabolism , Neurodegenerative Diseases/pathology , Protein Aggregation, Pathological/metabolism , Animals , Amyloidogenic Proteins/metabolism , Amyloidogenic Proteins/chemistry , Protein Aggregates , Alzheimer Disease/metabolism , Alzheimer Disease/pathology , Phase Separation
10.
J Environ Manage ; 351: 119839, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38104464

ABSTRACT

Photo-enhanced Biological Phosphorus Removal (PEBPR) systems, promising wastewater treatment technology, offer efficient phosphorus removal without external oxygen. However, comprehending the impact of sludge retention time (SRT) on the system is crucial for successful implementation. This study investigated the SRT effect on nutrient fate, microbial community, and bacterial phototolerance in PEBPR systems. PEBPR systems exhibited good bacterial phototolerance at SRT of 10, 15, and 20Ā d, with optimal phosphorus-accumulation metabolism observed at SRT of 10 and 15d. However, at SRT of 5d, increased light sensitivity and glycogen-accumulating organisms (GAOs) growth resulted in poor P removal (71.9%). Accumulibacter-IIC were the dominant P accumulating organisms (PAOs) at SRT of 10, 15, and 20Ā d. Accumulibacter-I, IIC and IIF were the major PAOs at SRT of 5Ā d. The decrease in SRT promoted the microalgal population diversity, and Dictyosphaerium and Chlorella were the major microalgal species in this study. Flow cytometry results revealed high light intensity triggered intracellular Fe2+ efflux, limiting translation activity and metabolism. Moreover, PAOs had lower phototolerance than GAOs due to Poly-P bound intracellular Mg2+ affecting enzyme activity. This study provides an in-depth understanding of PEBPR systems operation strategy toward environmentally sustainable wastewater treatment.


Subject(s)
Chlorella , Microbiota , Sewage , Phosphorus/metabolism , Bioreactors/microbiology , Bacteria/metabolism , Nutrients
11.
Molecules ; 29(5)2024 Feb 21.
Article in English | MEDLINE | ID: mdl-38474452

ABSTRACT

Cynomorium songaricum Rupr. (CSR) belongs to the family Cynomoriaceae. It is a perennial succulent parasitic herb with a reddish-brown coloration, predominantly submerged in sand and lacking chlorophyll. Traditionally, it has been used in ethnic medicine to treat various diseases, such as gastric ulcers, indigestion, bowel movements, and improving sexual function. To comprehensively collect CSR data, extensive literature searches were conducted using medical, ecological, and scientific databases such as Google Scholar, PubMed, Science Direct, Web of Science, and China National Knowledge Infrastructure (CNKI). This article summarizes and categorizes research on the uses, phytochemical characteristics, pharmacological activities, and toxicity of ethnic medicine, with the aim of establishing a solid foundation and proposing new avenues for exploring and developing potential applications of CSR. So far, a total of 98 compounds have been isolated and identified from CSR, including flavonoids, terpenes, steroids, and other compounds. It is worth noting that flavonoids and polysaccharides have significant antioxidant and anti-inflammatory properties. In addition, these compounds also show good application prospects in anti-tumor, antioxidant, anti-aging, anti-fatigue, anti-diabetes, and other aspects. Although extensive progress has been made in the basic research of CSR, further research is still needed to enhance the understanding of its mechanism of action and explore more unknown compounds. Our review indicates that CSR has broad prospects and deserves further research.


Subject(s)
Cynomorium , Ethnopharmacology , Antioxidants , Medicine, Chinese Traditional , Phytochemicals/pharmacology , Flavonoids , Plant Extracts/chemistry , Phytotherapy
12.
Molecules ; 29(12)2024 Jun 12.
Article in English | MEDLINE | ID: mdl-38930866

ABSTRACT

The H5N1 avian influenza virus seriously affects the health of poultry and humans. Once infected, the mortality rate is very high. Therefore, accurate and timely detection of the H5N1 avian influenza virus is beneficial for controlling its spread. This article establishes a dual gene detection method based on dual RPA for simultaneously detecting the HA and M2 genes of H5N1 avian influenza virus, for the detection of H5N1 avian influenza virus. Design specific primers for the conserved regions of the HA and M2 genes. The sensitivity of the dual RT-RPA detection method for HA and M2 genes is 1 Ɨ 10-7 ng/ĀµL. The optimal primer ratio is 1:1, the optimal reaction temperature is 40 Ā°C, and the optimal reaction time is 20 min. Dual RT-RPA was used to detect 72 samples, and compared with RT-qPCR detection, the Kappa value was 1 (p value < 0.05), and the clinical sample detection sensitivity and specificity were both 100%. The dual RT-RPA method is used for the first time to simultaneously detect two genes of the H5N1 avian influenza virus. As an accurate and convenient diagnostic tool, it can be used to diagnose the H5N1 avian influenza virus.


Subject(s)
Influenza A Virus, H5N1 Subtype , Influenza in Birds , Influenza A Virus, H5N1 Subtype/genetics , Animals , Influenza in Birds/virology , Influenza in Birds/diagnosis , Humans , Sensitivity and Specificity , Influenza, Human/virology , Influenza, Human/diagnosis , Viral Matrix Proteins/genetics , Hemagglutinin Glycoproteins, Influenza Virus/genetics , Birds/virology , Viroporin Proteins
13.
J Integr Plant Biol ; 66(4): 709-730, 2024 Apr.
Article in English | MEDLINE | ID: mdl-38483018

ABSTRACT

Hybrid rice (Oryza sativa) generally outperforms its inbred parents in yield and stress tolerance, a phenomenon termed heterosis, but the underlying mechanism is not completely understood. Here, we combined transcriptome, proteome, physiological, and heterosis analyses to examine the salt response of super hybrid rice Chaoyou1000 (CY1000). In addition to surpassing the mean values for its two parents (mid-parent heterosis), CY1000 exhibited a higher reactive oxygen species scavenging ability than both its parents (over-parent heterosis or heterobeltiosis). Nonadditive expression and allele-specific gene expression assays showed that the glutathione S-transferase gene OsGSTU26 and the amino acid transporter gene OsAAT30 may have major roles in heterosis for salt tolerance, acting in an overdominant fashion in CY1000. Furthermore, we identified OsWRKY72 as a common transcription factor that binds and regulates OsGSTU26 and OsAAT30. The salt-sensitive phenotypes were associated with the OsWRKY72paternal genotype or the OsAAT30maternal genotype in core rice germplasm varieties. OsWRKY72paternal specifically repressed the expression of OsGSTU26 under salt stress, leading to salinity sensitivity, while OsWRKY72maternal specifically repressed OsAAT30, resulting in salinity tolerance. These results suggest that the OsWRKY72-OsAAT30/OsGSTU26 module may play an important role in heterosis for salt tolerance in an overdominant fashion in CY1000 hybrid rice, providing valuable clues to elucidate the mechanism of heterosis for salinity tolerance in hybrid rice.


Subject(s)
Hybrid Vigor , Oryza , Hybrid Vigor/genetics , Reactive Oxygen Species/metabolism , Oryza/genetics , Oryza/metabolism , Salt Tolerance/genetics , Phenotype
14.
J Biol Chem ; 298(3): 101731, 2022 03.
Article in English | MEDLINE | ID: mdl-35176281

ABSTRACT

Lung cancer has the highest mortality among cancers worldwide due to its high incidence and lack of the effective cures. We have previously demonstrated that the membrane ion channel TMEM16A is a potential drug target for the treatment of lung adenocarcinoma and have identified a pocket of inhibitor binding that provides the basis for screening promising new inhibitors. However, conventional drug discovery strategies are lengthy and costly, and the unpredictable side effects lead to a high failure rate in drug development. Therefore, finding new therapeutic directions for already marketed drugs may be a feasible strategy to obtain safe and effective therapeutic drugs. Here, we screened a library of over 1400 Food and Drug Administration-approved drugs through virtual screening and activity testing. We identified a drug candidate, Zafirlukast (ZAF), clinically approved for the treatment of asthma, that could inhibit the TMEM16A channel in a concentration-dependent manner. Molecular dynamics simulations and site-directed mutagenesis experiments showed that ZAF can bind to S387/N533/R535 in the nonselective inhibitor binding pocket, thereby blocking the channel pore. Furthermore, we demonstrate ZAF can target TMEM16A channel to inhibit the proliferation and migration of lung adenocarcinoma LA795Ā cells. InĀ vivo experiments showed that ZAF can significantly inhibit lung adenocarcinoma tumor growth in mice. Taken together, we identified ZAF as a novel TMEM16A channel inhibitor with excellent anticancer activity, and as such, it represents a promising candidate for future preclinical and clinical studies.


Subject(s)
Adenocarcinoma of Lung , Anoctamin-1 , Indoles , Lung Neoplasms , Phenylcarbamates , Sulfonamides , Adenocarcinoma of Lung/drug therapy , Adenocarcinoma of Lung/genetics , Adenocarcinoma of Lung/metabolism , Animals , Anoctamin-1/antagonists & inhibitors , Anoctamin-1/metabolism , Chloride Channels , Indoles/pharmacology , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Lung Neoplasms/metabolism , Mice , Phenylcarbamates/pharmacology , Sulfonamides/pharmacology
15.
Plant Cell ; 32(5): 1626-1643, 2020 05.
Article in English | MEDLINE | ID: mdl-32184349

ABSTRACT

Ethylene plays important roles in plant growth and development, but the regulation of ethylene signaling is largely unclear, especially in crops such as rice (Oryza sativa). Here, by analysis of the ethylene-insensitive mutant mao huzi 11 (mhz11), we identified the GDSL lipase MHZ11, which modulates ethylene signaling in rice roots. MHZ11 localized to the endoplasmic reticulum membrane and has acyl-hydrolyzing activity. This activity affects the homeostasis of sterols in rice roots and is required for root ethylene response. MHZ11 overexpression caused constitutive ethylene response in roots. Genetically, MHZ11 acts with the ethylene receptor ETHYLENE RESPONSE SENSOR2 (OsERS2) upstream of CONSTITUTIVE TRIPLE RESPONSE2 (OsCTR2) and ETHYLENE INSENSITIVE2 (OsEIN2). The mhz11 mutant maintains more OsCTR2 in the phosphorylated form whereas MHZ11 overexpression promotes ethylene-mediated inhibition of OsCTR2 phosphorylation. MHZ11 colocalized with the ethylene receptor OsERS2, and its effect on OsCTR2 phosphorylation requires ethylene perception and initiation of ethylene signaling. The mhz11 mutant overaccumulated sterols and blocking sterol biosynthesis partially rescued the mhz11 ethylene response, likely by reducing receptor-OsCTR2 interaction and OsCTR2 phosphorylation. We propose that MHZ11 reduces sterol levels to impair receptor-OsCTR2 interactions and OsCTR2 phosphorylation for triggering ethylene signaling. Our study reveals a mechanism by which MHZ11 participates in ethylene signaling for regulation of root growth in rice.


Subject(s)
Ethylenes/metabolism , Lipase/metabolism , Oryza/metabolism , Plant Roots/metabolism , Signal Transduction , Endoplasmic Reticulum/metabolism , Genes, Plant , Hydrolysis , Lipid Metabolism , Mutation/genetics , Oryza/genetics , Phenotype , Phosphorylation , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Roots/genetics , Plants, Genetically Modified
16.
Eur J Clin Invest ; 53(12): e14067, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37515404

ABSTRACT

BACKGROUND: Observational studies have suggested an association between lipid-lowering drugs and inflammatory bowel disease (IBD) risk. This study aimed to assess the causal influence of lipid-lowering agents on IBD risk using Mendelian randomization analysis. METHOD: In a population of 173,082 individuals of European ancestry, 55 single-nucleotide polymorphisms were identified as instrumental variables for 6 lipid-lowering drug targets (HMGCR, NPC1LC, PCSK9, LDLR, CETP and APOB). Summary statistics for the genome-wide association study of IBD, ulcerative colitis (UC) and Crohn's disease (CD) were obtained from the FinnGen consortium, Program in Complex Trait Genomics and UK Biobank. Inverse-variance weighted was employed as the primary MR method, and odds ratios (ORs) with 95% confidence intervals were reported as the results. Sensitivity analyses using conventional MR methods were conducted to assess result robustness. RESULTS: Gene-proxied inhibition of Niemann-Pick C1-like 1 (NPC1L1) was associated with an increased IBD risk (OR [95% CI]: 2.31 [1.38, 3.85]; p = .001), particularly in UC (OR [95% CI]: 2.40 [1.21, 4.74], p = .012), but not in CD. This finding was replicated in the validation cohort. Additionally, gene-proxied inhibition of low-density lipoprotein receptor was associated with reduced IBD (OR [95% CI]: .72 [.60, .87], p < .001) and UC risk (OR [95% CI]: .74 [.59, .92], p = .006), although this result was not replicated in the validation cohort. Other drug targets did not show significant associations with IBD, UC or CD risk. CONCLUSION: Inhibition of the lipid-lowering drug-target NPC1L1 leads to an increased IBD risk, mainly in the UC population.


Subject(s)
Colitis, Ulcerative , Crohn Disease , Inflammatory Bowel Diseases , Humans , Proprotein Convertase 9 , Genome-Wide Association Study , Inflammatory Bowel Diseases/drug therapy , Inflammatory Bowel Diseases/epidemiology , Inflammatory Bowel Diseases/genetics , Colitis, Ulcerative/drug therapy , Colitis, Ulcerative/epidemiology , Colitis, Ulcerative/genetics , Crohn Disease/drug therapy , Crohn Disease/epidemiology , Crohn Disease/genetics , Hypolipidemic Agents , Lipids
17.
Org Biomol Chem ; 21(47): 9422-9427, 2023 Dec 06.
Article in English | MEDLINE | ID: mdl-37987684

ABSTRACT

Enol ethers are essential synthetic frameworks and widely applied in organic synthesis; however, high regio- and stereo-selective access to enol ethers remains challenging. Herein, we report a titanium-catalyzed stereospecific anti-Markovnikov hydroalkoxylation reaction of alkynes for the synthesis of Z-enol ethers with excellent functional group tolerance and yields. Mechanistic studies showed that the titanium coordinates with the alkyne and then an oxygen anion attacks the π-bond of the alkyne from the backside to provide a trans-oxygen metallation intermediate, which accounts for the high Z-stereoselectivity. Furthermore, Z-enol ethers could be applied as a kind of synthon for late-stage transformations and gram-scale synthesis, which demonstrates their potential value in organic synthesis.

18.
Nucleic Acids Res ; 49(10): 5451-5469, 2021 06 04.
Article in English | MEDLINE | ID: mdl-33836076

ABSTRACT

Deoxyribonucleic acid (DNA) has evolved to be a naturally selected, robust biomacromolecule for gene information storage, and biological evolution and various diseases can find their origin in uncertainties in DNA-related processes (e.g. replication and expression). Recently, synthetic DNA has emerged as a compelling molecular media for digital data storage, and it is superior to the conventional electronic memory devices in theoretical retention time, power consumption, storage density, and so forth. However, uncertainties in the in vitro DNA synthesis and sequencing, along with its conjugation chemistry and preservation conditions can lead to severe errors and data loss, which limit its practical application. To maintain data integrity, complicated error correction algorithms and substantial data redundancy are usually required, which can significantly limit the efficiency and scale-up of the technology. Herein, we summarize the general procedures of the state-of-the-art DNA-based digital data storage methods (e.g. write, read, and preservation), highlighting the uncertainties involved in each step as well as potential approaches to correct them. We also discuss challenges yet to overcome and research trends in the promising field of DNA-based data storage.


Subject(s)
DNA , Genes, Synthetic , Information Storage and Retrieval/methods , DNA/chemical synthesis , DNA/chemistry
19.
Proc Natl Acad Sci U S A ; 117(38): 23499-23509, 2020 09 22.
Article in English | MEDLINE | ID: mdl-32907946

ABSTRACT

Understanding the molecular basis of male sterility and developing practical male-sterility systems are essential for heterosis utilization and commercial hybrid seed production in crops. Here, we report molecular regulation by genic male-sterility gene maize male sterility 7 (ZmMs7) and its application for developing a dominant male-sterility system in multiple species. ZmMs7 is specifically expressed in maize anthers, encodes a plant homeodomain (PHD) finger protein that functions as a transcriptional activator, and plays a key role in tapetal development and pollen exine formation. ZmMs7 can interact with maize nuclear factor Y (NF-Y) subunits to form ZmMs7-NF-YA6-YB2-YC9/12/15 protein complexes that activate target genes by directly binding to CCAAT box in their promoter regions. Premature expression of ZmMs7 in maize by an anther-specific promoter p5126 results in dominant and complete male sterility but normal vegetative growth and female fertility. Early expression of ZmMs7 downstream genes induced by prematurely expressed ZmMs7 leads to abnormal tapetal development and pollen exine formation in p5126-ZmMs7 maize lines. The p5126-ZmMs7 transgenic rice and Arabidopsis plants display similar dominant male sterility. Meanwhile, the mCherry gene coupled with p5126-ZmMs7 facilitates the sorting of dominant sterility seeds based on fluorescent selection. In addition, both the ms7-6007 recessive male-sterility line and p5126-ZmMs7M dominant male-sterility line are highly stable under different genetic germplasms and thus applicable for hybrid maize breeding. Together, our work provides insight into the mechanisms of anther and pollen development and a promising technology for hybrid seed production in crops.


Subject(s)
Gene Expression Regulation, Plant/genetics , Plant Infertility/genetics , Plant Proteins/genetics , Promoter Regions, Genetic/genetics , Zea mays/genetics , Arabidopsis/genetics , Crops, Agricultural , Oryza/genetics , Plants, Genetically Modified/genetics , Plants, Genetically Modified/growth & development , Pollen/genetics , Zea mays/growth & development
20.
Int J Mol Sci ; 24(19)2023 Oct 08.
Article in English | MEDLINE | ID: mdl-37834437

ABSTRACT

The recent pandemic of SARS-CoV-2 has underscored the critical need for rapid and precise viral detection technologies. Point-of-care (POC) technologies, which offer immediate and accurate testing at or near the site of patient care, have become a cornerstone of modern medicine. Prokaryotic Argonaute proteins (pAgo), proficient in recognizing target RNA or DNA with complementary sequences, have emerged as potential game-changers. pAgo present several advantages over the currently popular CRISPR/Cas systems-based POC diagnostics, including the absence of a PAM sequence requirement, the use of shorter nucleic acid molecules as guides, and a smaller protein size. This review provides a comprehensive overview of pAgo protein detection platforms and critically assesses their potential in the field of viral POC diagnostics. The objective is to catalyze further research and innovation in pAgo nucleic acid detection and diagnostics, ultimately facilitating the creation of enhanced diagnostic tools for clinic viral infections in POC settings.


Subject(s)
Nucleic Acids , Point-of-Care Systems , Humans , Argonaute Proteins/genetics , Argonaute Proteins/metabolism , Prokaryotic Cells/metabolism , Point-of-Care Testing , CRISPR-Cas Systems
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