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Genes Dec 2022Drought is a primary environmental factor limiting maize production globally. Although transferring a single gene to maize can enhance drought resistance, maize response...
Drought is a primary environmental factor limiting maize production globally. Although transferring a single gene to maize can enhance drought resistance, maize response to water deficit requires further improvement to accommodate the steadily intensifying drought events worldwide. Here, we generated dual transgene lines simultaneously overexpressing two drought-resistant genes, (encoding a vacuolar-type H+ pyrophosphatase) and (encoding a NAM, ATAF, and CUC (NAC)-type transcription factor). Following drought stress, survival rates of the pyramided transgenic seedlings reached 62-66%, while wild-type and single transgene seedling survival rates were 23% and 37-42%, respectively. Maize seedlings co-expressing and exhibited higher photosynthesis rates, antioxidant enzyme activities, and root-shoot ratios than the wild type, and anthesis-silking intervals were shorter while grain yields were higher under water deficit conditions in field trials. Additionally, RNA-sequencing analysis confirmed that photosynthesis and stress-related metabolic processes were stimulated in the dual transgene plants under drought conditions. The findings in this work illustrate how high co-expression of different drought-related genes can reinforce drought resistance over that of individual transgene lines, providing a path for developing arid climate-adapted elite maize varieties.
Topics: Drought Resistance; Zea mays; Plants, Genetically Modified; Seedlings; Water
PubMed: 36672748
DOI: 10.3390/genes14010008 -
International Journal of General... 2021ENPP1 and ENTPD1 are two main enzymes involved in ATP-AMP-ADP-adenosine axis, which is associated with lipid metabolism, diabetes mellitus (DM) and renal fibrosis. The...
INTRODUCTION
ENPP1 and ENTPD1 are two main enzymes involved in ATP-AMP-ADP-adenosine axis, which is associated with lipid metabolism, diabetes mellitus (DM) and renal fibrosis. The single nucleotide polymorphisms (SNPs) of ENPP1 and ENTPD1, rs1044498 and rs6584026, are associated with these factors. This retrospective study aimed to address the two SNPs variants in hemodialysis (HD) patients and analyzes their relations with clinical characteristics.
METHODS
This study included 543 regular HD patients over 3 months at our center. Overnight fasting peripheral blood sample was taken from each subject to extract the DNA. The genotypes of rs1044498 and rs6584026 were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The basic clinical data were noted such as sex, age, and HD-age, and the main causes of chronic kidney disease (CKD) and the clinical characteristics were collected on average at least three times in half a year. -test and Chi-test were performed for the statistical analyses. Binary logistic regression was applied for the significant parameters by excluding the confounders, gender, age and HD-age. All statistical tests were considered significant for <0.05.
RESULTS
The rs1044498 genotypes showed in two types, A/A and A/C without C/C. The rs6584026 genotypes were C/C and C/T without T/T. The genotype frequency of rs1044498 (A/C) was 0.238, and the genotype frequency of rs6584026 (C/T) was 0.328. The age and the level of lipoprotein α showed statistical significance with rs1044498 variant (A/C, <0.05). The rs6584026 variant (C/T) was frequently found in patients with nephritis (<0.05). The albumin, alkaline phosphatase (ALP), lipoprotein α, cholesterol, apolipoprotein B (Apo B), Apo B/A1 and nephritis were independently associated with rs6584026 variant (C/T, <0.05) in binary logistic regression model by controlling the confounders of gender, age and HD-age. High level of triglyceride and low level of urine nitrogen were related to rs6584026 variant (C/T, <0.05).
CONCLUSION
The rs1044498 and rs6584026 SNPs were related to several high levels of lipids, and rs6584026 variant was related to nephritis and autoimmune disease. The rs6584026 SNP may contribute to the increased risks of cholesterol and ApoB/A1 in HD patients.
PubMed: 34675608
DOI: 10.2147/IJGM.S332911 -
Current Opinion in Structural Biology Dec 2022DNA polymerases play central roles in DNA replication and repair by catalyzing template-directed nucleotide incorporation. Recently time-lapse X-ray crystallography,... (Review)
Review
DNA polymerases play central roles in DNA replication and repair by catalyzing template-directed nucleotide incorporation. Recently time-lapse X-ray crystallography, which allows one to observe reaction intermediates, has revealed numerous and unexpected mechanistic features of DNA polymerases. In this article, we will examine recent new discoveries that have come from time-lapse crystallography that are currently transforming our understanding of the structural mechanisms used by DNA polymerases. Among these new discoveries are the binding of a third metal ion within the polymerase active site, the mechanisms of translocation along the DNA, the presence of new fidelity checkpoints, a novel pyrophosphatase activity within the active site, and the mechanisms of pyrophosphorolysis.
Topics: Time-Lapse Imaging; DNA-Directed DNA Polymerase; DNA; Crystallography, X-Ray; DNA Repair; DNA Replication
PubMed: 36174287
DOI: 10.1016/j.sbi.2022.102465 -
Frontiers in Molecular Biosciences 2019Membrane integral pyrophosphatases (mPPases) are responsible for the hydrolysis of pyrophosphate. This enzymatic mechanism is coupled to the pumping of H or Na across... (Review)
Review
Membrane integral pyrophosphatases (mPPases) are responsible for the hydrolysis of pyrophosphate. This enzymatic mechanism is coupled to the pumping of H or Na across membranes in a process that can be K dependent or independent. Understanding the movements and dynamics throughout the mPPase catalytic cycle is important, as this knowledge is essential for improving or impeding protein function. mPPases have been shown to play a crucial role in plant maturation and abiotic stress tolerance, and so have the potential to be engineered to improve plant survival, with implications for global food security. mPPases are also selectively toxic drug targets, which could be pharmacologically modulated to reduce the virulence of common human pathogens. The last few years have seen the publication of many new insights into the function and structure of mPPases. In particular, there is a new body of evidence that the catalytic cycle is more complex than originally proposed. There are structural and functional data supporting a mechanism involving half-of-the-sites reactivity, inter-subunit communication, and exit channel motions. A more advanced and in-depth understanding of mPPases has begun to be uncovered, leaving the field of research with multiple interesting avenues for further exploration and investigation.
PubMed: 31824962
DOI: 10.3389/fmolb.2019.00132 -
Nature Chemical Biology Oct 2020The NUDIX hydrolase NUDT15 was originally implicated in sanitizing oxidized nucleotides, but was later shown to hydrolyze the active thiopurine metabolites,...
The NUDIX hydrolase NUDT15 was originally implicated in sanitizing oxidized nucleotides, but was later shown to hydrolyze the active thiopurine metabolites, 6-thio-(d)GTP, thereby dictating the clinical response of this standard-of-care treatment for leukemia and inflammatory diseases. Nonetheless, its physiological roles remain elusive. Here, we sought to develop small-molecule NUDT15 inhibitors to elucidate its biological functions and potentially to improve NUDT15-dependent chemotherapeutics. Lead compound TH1760 demonstrated low-nanomolar biochemical potency through direct and specific binding into the NUDT15 catalytic pocket and engaged cellular NUDT15 in the low-micromolar range. We also employed thiopurine potentiation as a proxy functional readout and demonstrated that TH1760 sensitized cells to 6-thioguanine through enhanced accumulation of 6-thio-(d)GTP in nucleic acids. A biochemically validated, inactive structural analog, TH7285, confirmed that increased thiopurine toxicity takes place via direct NUDT15 inhibition. In conclusion, TH1760 represents the first chemical probe for interrogating NUDT15 biology and potential therapeutic avenues.
Topics: Binding Sites; Cell Line; Drug Design; Drug Development; Escherichia coli; Humans; Inorganic Pyrophosphatase; Models, Molecular; Protein Binding; Protein Conformation; Pyrophosphatases; Structure-Activity Relationship
PubMed: 32690945
DOI: 10.1038/s41589-020-0592-z -
Cell Death & Disease Feb 2023Impaired protein N-glycosylation leads to the endoplasmic reticulum (ER) stress, which triggers adaptive survival or maladaptive apoptosis in renal tubules in diabetic...
Impaired protein N-glycosylation leads to the endoplasmic reticulum (ER) stress, which triggers adaptive survival or maladaptive apoptosis in renal tubules in diabetic kidney disease (DKD). Therapeutic strategies targeting ER stress are promising for the treatment of DKD. Here, we report a previously unappreciated role played by ENTPD5 in alleviating renal injury by mediating ER stress. We found that ENTPD5 was highly expressed in normal renal tubules; however, ENTPD5 was dynamically expressed in the kidney and closely related to pathological DKD progression in both human patients and mouse models. Overexpression of ENTPD5 relieved ER stress in renal tubular cells, leading to compensatory cell proliferation that resulted in hypertrophy, while ENTPD5 knockdown aggravated ER stress to induce cell apoptosis, leading to renal tubular atrophy and interstitial fibrosis. Mechanistically, ENTPD5-regulated N-glycosylation of proteins in the ER to promote cell proliferation in the early stage of DKD, and continuous hyperglycemia activated the hexosamine biosynthesis pathway (HBP) to increase the level of UDP-GlcNAc, which driving a feedback mechanism that inhibited transcription factor SP1 activity to downregulate ENTPD5 expression in the late stage of DKD. This study was the first to demonstrate that ENTPD5 regulated renal tubule cell numbers through adaptive proliferation or apoptosis in the kidney by modulating the protein N-glycosylation rate in the ER, suggesting that ENTPD5 drives cell fate in response to metabolic stress and is a potential therapeutic target for renal diseases.
Topics: Animals; Humans; Mice; Glycosylation; Kidney; Kidney Tubules; Oncogene Proteins; Pyrophosphatases; Endoplasmic Reticulum Stress
PubMed: 36849424
DOI: 10.1038/s41419-023-05685-4 -
Communications Biology Nov 2020Guanosine 3',5'-bis(pyrophosphate) (ppGpp) functions as a second messenger in bacteria to adjust their physiology in response to environmental changes. In recent years,...
Guanosine 3',5'-bis(pyrophosphate) (ppGpp) functions as a second messenger in bacteria to adjust their physiology in response to environmental changes. In recent years, the ppGpp-specific hydrolase, metazoan SpoT homolog-1 (Mesh1), was shown to have important roles for growth under nutrient deficiency in Drosophila melanogaster. Curiously, however, ppGpp has never been detected in animal cells, and therefore the physiological relevance of this molecule, if any, in metazoans has not been established. Here, we report the detection of ppGpp in Drosophila and human cells and demonstrate that ppGpp accumulation induces metabolic changes, cell death, and eventually lethality in Drosophila. Our results provide the evidence of the existence and function of the ppGpp-dependent stringent response in animals.
Topics: Animals; Bacteria; Drosophila melanogaster; Guanosine Pentaphosphate; Guanosine Tetraphosphate; Pyrophosphatases; Second Messenger Systems; Signal Transduction
PubMed: 33188280
DOI: 10.1038/s42003-020-01368-4 -
Frontiers in Plant Science 2023This research aimed to assess the feasibility of utilizing Raman spectroscopy in plant breeding programs. For this purpose, the evaluation of the mutant populations set...
This research aimed to assess the feasibility of utilizing Raman spectroscopy in plant breeding programs. For this purpose, the evaluation of the mutant populations set up the application of 4 mM NaN to the somatic embryos obtained from mature wheat ( L. Adana-99 cv.) embryos. Advanced wheat mutant lines, which were brought up to the seventh generation with salt stress tolerance by following and environments constructed by mutated populations, were evaluated using conventional techniques [measurement of antioxidant enzyme activities (SOD, CAT, and POX), total chlorophyll, TBARS, and proline contents; measurement of the concentration of Na and K ions; and evaluation of gene expression by qPCR (HKT2;1, HKT1;5, SOS1, Na/H vacuolar antiporter, V-PPase, V-ATPase, and P5CS)] and Raman spectroscopy. In this research, no significant difference was found in the increase of SOD, CAT, and POX antioxidant enzyme activities between the salt-treated and untreated experimental groups of the commercial cultivar, while there was a statistically significant increase in salt-treated advanced generation mutant lines as compared to control and the salt-treated commercial cultivar. Proline showed a statistically significant increase in all experimental groups compared to the untreated commercial cultivar. The degradation in the amount of chlorophyll was lower in the salt-treated advanced generation mutant lines than in the salt-treated commercial cultivar. According to gene expression studies, there were statistical differences at various levels in terms of Na and/or K uptake from soil to plant (THKT2;1, HKT1;5, and SOS1), and Na compartmentalizes into the cell vacuole (Na/H vacuolar antiporter, vacuolar pyrophosphatase, and vacuola H-ATPase). The expression activity of P5CS, which is responsible for the transcription of proline, is similar to the content of proline in the current study. As a result of Raman spectroscopy, the differences in peaks represent the protein-related bands in mutant lines having a general decreasing trend in intensity when compared to the commercial cultivar. Amide-I (1,630 and 1,668 cm), Histidine, Lysine, Arginine, and Leucine bands (823, 849, 1,241, 1,443, and 1,582 cm) showed decreasing wavenumbers. Beta-carotene peaks at 1,153 and 1,519 cm showed increasing trends when the normalized Raman intensities of the mutant lines were compared.
PubMed: 36909443
DOI: 10.3389/fpls.2023.1116876 -
Heliyon Jan 2024The need for naturally occurring constituents is driven by the rise in the cancer prevalence and the unpleasant side effects associated with chemotherapeutics.... (Review)
Review
The need for naturally occurring constituents is driven by the rise in the cancer prevalence and the unpleasant side effects associated with chemotherapeutics. Triptolide, the primary active component of "Tripterygium Wilfordii", has exploited for biological mechanisms and therapeutic potential against various tumors. Based on the recent pre-clinical investigations, triptolide is linked to the induction of death of cancerous cells by triggering cellular apoptosis via inhibiting heat shock protein expression (HSP70), and cyclin dependent kinase (CDKs) by up regulating expression of P21. MKP1, histone methyl transferases and RNA polymerases have all recently identified as potential targets of triptolide in cells. Autophagy, AKT signaling pathway and various pathways involving targeted proteins such as A-disintegrin & metalloprotease-10 (ADAM10), Polycystin-2 (PC-2), dCTP pyro-phosphatase 1 (DCTP1), peroxiredoxin-I (Prx-I), TAK1 binding protein (TAB1), kinase subunit (DNA-PKcs) and the xeroderma-pigmentosum B (XPB or ERCC3) have been exploited. Besides that, triptolide is responsible for enhancing the effectiveness of various chemotherapeutics. In addition, several triptolide moieties, including minnelide and LLDT8, have progressed in investigations on humans for the treatment of cancer. Targeted strategies, such as triptolide conjugation with ligands or triptolide loaded nano-carriers, are efficient techniques to confront toxicities associated with triptolide. We expect and anticipate that advances in near future, regarding combination therapies of triptolide, might be beneficial against cancerous cells.
PubMed: 38293343
DOI: 10.1016/j.heliyon.2024.e24335 -
Current Osteoporosis Reports Jun 2020This review summarizes current understanding of generalized arterial calcification of infancy (GACI), emphasizing pathophysiology, clinical presentation, and approaches... (Review)
Review
PURPOSE OF REVIEW
This review summarizes current understanding of generalized arterial calcification of infancy (GACI), emphasizing pathophysiology, clinical presentation, and approaches and controversies in management.
RECENT FINDINGS
Identification of causative ENPP1 mutations revealed that GACI arises from deficiencies in inorganic pyrophosphate (leading to calcifications) and adenosine monophosphate (leading to intimal proliferation). Identification of genotypic and phenotypic overlap with pseudoxanthoma elasticum and autosomal recessive hypophosphatemic rickets further advanced understanding of GACI as a complex, multisystemic disease. Clinical data is limited to small, retrospective samples; it is therefore unknown whether commonly used medications, such as bisphosphonates and hypophosphatemia treatment, are therapeutic or potentially harmful. ENPP1-Fc replacement represents a promising approach warranting further study. Knowledge gaps in natural history place clinicians at high risk of assigning causality to interventions that are correlated with changes in clinical status. There is thus a critical need for improved natural history studies to develop and test targeted therapies.
Topics: Adenosine Monophosphate; Bone Density Conservation Agents; Calcinosis; Cardiovascular Agents; Chelating Agents; Diphosphates; Diphosphonates; Familial Hypophosphatemic Rickets; Genotype; Hearing Loss; Humans; Multidrug Resistance-Associated Proteins; Phenotype; Phosphoric Diester Hydrolases; Pseudoxanthoma Elasticum; Pyrophosphatases; Thiosulfates; Tooth Diseases; Vascular Calcification; Vitamin D
PubMed: 32172442
DOI: 10.1007/s11914-020-00577-4