RESUMO
Protein adenosine diphosphate-ribosylation (ADPr) is caused by the covalent binding of one or more ADP-ribose moieties to a target protein and regulates the biological functions of the target protein. To fully understand the regulatory mechanism of ADP-ribosylation, the essential step is the identification of the ADPr sites from the proteome. As the experimental approaches are costly and time-consuming, it is necessary to develop a computational tool to predict ADPr sites. Recently, serine has been found to be the major residue type for ADP-ribosylation but no predictor is available. In this study, we collected thousands of experimentally validated human ADPr sites on serine residue and constructed several different machine-learning classifiers. We found that the hybrid model, dubbed DeepSADPr, which integrated the one-dimensional convolutional neural network (CNN) with the One-Hot encoding approach and the word-embedding approach, compared favourably to other models in terms of both ten-fold cross-validation and independent test. Its AUC values reached 0.935 for ten-fold cross-validation. Its values of sensitivity, accuracy and Matthews's correlation coefficient reached 0.933, 0.867 and 0.740, respectively, with the fixed specificity value of 0.80. Overall, DeepSADPr is the first classifier for predicting Serine ADPr sites, which is available at http://www.bioinfogo.org/DeepSADPr.
Assuntos
Processamento de Proteína Pós-Traducional , Serina , ADP-Ribosilação , Adenosina Difosfato Ribose/química , Adenosina Difosfato Ribose/metabolismo , Humanos , Proteoma , Serina/metabolismoRESUMO
N is essential for plant architecture, particularly tillering. However, whether and how N mediates panicle branching and influences rice grain yield remains unclear. In order to identify genes and pathways associated with N-regulated panicle branching, we treated rice with different concentrations of N to determine the key genes by transcriptomic analysis and function verification. We measured panicle growth in response to N, and found that panicle branching benefits from 2 mM exogenous N, and 2-5 mM N is essential for vascular bundle, phloem, and xylem development in these branches. Interestingly, total N concentrations increased continuously with N 0-2 mM and decreased continuously with N 5-15 mM, whereas the concentrations of amino acids Tyr and Val increased continuously with N 0-15 mM in the panicle. Furthermore, N metabolism, phytohormone signal transduction, stress response, and photosynthesis pathways play important roles in response to nitrogen of regulating panicle branching. Altered expression of key N-response amino acid transporter gene OsAAP15 positively regulated panicle branching at low N concentrations, however, OsAAP15 negatively influenced it at high N concentrations. Overexpression of OsAAP15 in the field significantly increased primary and secondary branches, filled grain number, and grain yield by regulating the concentrations of amino acids Tyr and Val in the panicle. Taken together, OsAAP15, an amino acid transporter in response to nitrogen concentration, could mediate panicle branching and grain yield, and it may have applications in rice breeding to improve grain yield under extreme N concentrations.
Assuntos
Oryza , Oryza/metabolismo , Melhoramento Vegetal , Grão Comestível/genética , Grão Comestível/metabolismo , Nitrogênio/metabolismoRESUMO
N6methyladenosine (m6A) serves an essential role in RNA modulation and is implicated in multiple malignancies, including colorectal cancer (CRC). Methyltransferaselike 3 (METTL3) is an important writer in m6A modification, however its role in CRC in modifying small nucleolar RNA host gene 1 (SNHG1), an oncogenic long noncoding RNA, remains unclear. In the present study, METTL3 expression in CRC was assessed using online bioinformatics analysis, immunohistochemistry staining, western blotting, reverse transcription (RT)quantitative PCR (qPCR) and cell transfections. Cell proliferation, migration and invasion were determined using functional Cell Counting Kit8 (CCK8) and Transwell assays. SNHG1 expression in CRC was evaluated using online bioinformatics analysis and RTqPCR. Methylated RNA immunoprecipitation qPCR was performed to assess m6A modification changes of SNHG1 mRNA. The present study demonstrated that METTL3 is upregulated in CRC tissues and cell lines. Moreover, METTL3 expression was associated with several unfavourable clinical features in patients with CRC, including the stage of lymph node metastases and overall survival. Functional Transwell and CCK8 assays demonstrated that knockdown of METTL3 suppressed CRC cell proliferation and migration. Furthermore, METTL3 was positively correlated with SNHG1 in CRC tissue, as indicated by analysis of data from The Cancer Genome Atlas. Mechanistically, SNHG1 contains 18 m6A modification sites. Through cell transfections and actinomycin D assays, the present study found that METTL3mediated m6A modification at these sites enhances the stability of SNHG1 in CRC cells. Finally, it was demonstrated that SNHG1 knockdown partially diminished the facilitative effect of METTL3 on CRC cell migration and proliferation. The present study concluded that METTL3, a potential biomarker for assessing overall survival and metastasis in CRC, may serve as an oncogene, promote SNHG1 m6A modification, improve the stability of SNHG1 and enhance SNHG1mediated oncogenic function in CRC.
Assuntos
Neoplasias Colorretais , Metiltransferases , RNA Longo não Codificante , Humanos , Linhagem Celular Tumoral , Proliferação de Células/genética , Neoplasias Colorretais/patologia , Metiltransferases/genética , Metiltransferases/metabolismo , RNA , RNA Longo não Codificante/genéticaRESUMO
Background: Radiotherapy plays a crucial role in the management of Cervical cancer (CC), as the development of resistance by cancer cells to radiotherapeutic interventions is a significant factor contributing to treatment failure in patients. However, the specific mechanisms that contribute to this resistance remain unclear. Currently, molecular targeted therapy, including mitochondrial genes, has emerged as a new approach in treating different types of cancers, gaining significant attention as an area of research in addressing the challenge of radiotherapy resistance in cancer. Methods: The present study employed a rigorous screening methodology within the TCGA database to identify a cohort of patients diagnosed with CC who had received radiotherapy treatment. The control group consisted of individuals who demonstrated disease stability or progression after undergoing radiotherapy. In contrast, the treatment group consisted of patients who experienced complete or partial remission following radiotherapy. Following this, we identified and examined the differentially expressed genes (DEGs) in the two cohorts. Subsequently, we conducted additional analyses to refine the set of excluded DEGs by employing the least absolute shrinkage and selection operator regression and random forest techniques. Additionally, a comprehensive analysis was conducted in order to evaluate the potential correlation between the expression of core genes and the extent of immune cell infiltration in patients diagnosed with CC. The mitochondrial-associated genes were obtained from the MITOCARTA 3.0. Finally, the verification of increased expression of the mitochondrial gene TMEM38A in individuals with CC exhibiting sensitivity to radiotherapy was conducted using reverse transcription quantitative polymerase chain reaction and immunohistochemistry assays. Results: This process ultimately led to the identification of 7 crucial genes, viz., GJA3, TMEM38A, ID4, CDHR1, SLC10A4, KCNG1, and HMGCS2, which were strongly associated with radiotherapy sensitivity. The enrichment analysis has unveiled a significant association between these 7 crucial genes and prominent signaling pathways, such as the p53 signaling pathway, KRAS signaling pathway, and PI3K/AKT/MTOR pathway. By utilizing these 7 core genes, an unsupervised clustering analysis was conducted on patients with CC, resulting in the categorization of patients into three distinct molecular subtypes. In addition, a predictive model for the sensitivity of CC radiotherapy was developed using a neural network approach, utilizing the expression levels of these 7 core genes. Moreover, the CellMiner database was utilized to predict drugs that are closely linked to these 7 core genes, which could potentially act as crucial agents in overcoming radiotherapy resistance in CC. Conclusion: To summarize, the genes GJA3, TMEM38A, ID4, CDHR1, SLC10A4, KCNG1, and HMGCS2 were found to be closely correlated with the sensitivity of CC to radiotherapy. Notably, TMEM38A, a mitochondrial gene, exhibited the highest degree of correlation, indicating its potential as a crucial biomarker for the modulation of radiotherapy sensitivity in CC.
Assuntos
Neoplasias do Colo do Útero , Feminino , Humanos , Algoritmos , Proteínas Relacionadas a Caderinas , Genes Mitocondriais , Marcadores Genéticos , Proteínas do Tecido Nervoso , Fosfatidilinositol 3-Quinases , Neoplasias do Colo do Útero/genética , Neoplasias do Colo do Útero/radioterapiaRESUMO
Melatonin plays an important role in plant resistance to biotic and abiotic stresses. However, whether melatonin is involved in the regulation of plant architecture, such as the formation of axillary bud outgrowth or tillering, in rice remains unknown. Here, we found that different concentrations of melatonin influenced axillary bud outgrowth in rice, and moderate melatonin concentrations also alleviated the inhibition of axillary bud outgrowth in the presence of high concentrations of basic amino acids lysine and arginine. Furthermore, transcriptome analysis demonstrated that genes involved in nitrogen metabolism and phytohormone signal transduction pathways may affect axillary bud outgrowth, which is regulated by melatonin. We determined that the differentially expressed genes glutamine synthetase OsGS2 and amino acid transporter OsAAP14, which are involved in nitrogen metabolism and are regulated by melatonin and basic amino acids, were the key regulators of axillary bud outgrowth in rice. In addition, we validated the functions of OsGS2 and OsAAP14 using rice transgenic plants with altered axillary bud outgrowth and tillers. Taken together, these results suggest that melatonin mediates axillary bud outgrowth by improving nitrogen assimilation and transport in rice.
RESUMO
BACKGROUND: Cyclin-dependent Kinase 9 as one of the serine/threonine protein kinases has become an important target for the treatment of cancer especially driven by transcriptional dysregulation. OBJECTIVE: This thesis was conducted to elucidate the structure-activity relationship and interaction mode of coumarin compounds acting on CDK9. METHODS: Three-dimensional quantitative structure-activity relationship (3D-QSAR), molecular docking and molecular dynamics simulation were conducted to reveal the structural requirements for bioactivities. The 3D-QSAR model was constructed to find the features required for different substituents on the coumarin scaffold. Molecular docking and molecular dynamics simulation were employed to generate the binding mode and stability of CDK9. RESULTS: The Q2 and R2 values of the CoMFA model were calculated as 0.52 and 0.999, while those for the CoMSIA model were 0.606 and 0.998. It is believed that the significant statistical parameters of CoMFA and CoMSIA models revealed high activity-descriptor relationship efficiency. Therefore, we considered the 3D-QSAR model to be robust and accurate. The contour maps provided a deep structure-activity relationship and valuable clues for rational modification. Based on the contour maps, 4 novel CDK9 inhibitors which were predicted to have satisfactory pharmacokinetic characteristics were designed and exhibited better-predicted activities. Subsequently, molecular docking was employed to generate the binding mode of CDK9. Furthermore, 50 ns MD simulation was of great help in verifying the accuracy of docking results and the stability of the complexes. CONCLUSION: The study is a valuable insight for further research on novel and effective inhibitors targeting CDK9.
Assuntos
Simulação de Dinâmica Molecular , Relação Quantitativa Estrutura-Atividade , Simulação de Acoplamento Molecular , Quinase 9 Dependente de Ciclina , Cumarínicos/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Inibidores de Proteínas Quinases/químicaRESUMO
BACKGROUND: More and more evidence indicates that microRNAs are present and involved in many tumor-related diseases. The function of microRNA-622 (miR-622) in colorectal cancer (CRC) remains controversial. Dual specificity tyrosine phosphorylation-regulated kinase 2 (DYRK2) has been reported as a tumor suppressor gene in different cancers. The detailed regulation mechanism of DYRK2 in CRC remains unclear. METHODS: miR-622 and DYRK2 expression levels were detected at tissue and cellular level respectively by using real time polymerase chain reaction (PCR), Western blot, and immunohistochemical staining. Pearson's correlation analysis was used to evaluate the correlation between miR-622 and DYRK2. Transwell assay was applied to measure the effect of miR-622 on migration and invasion of SW1116 and SW480. We used dual luciferase reporter assay to confirm the targeted binding effect of miR-622 and DYRK2 3'-untranslated region (3'UTR). An antisense experiment was executed to further confirm the role miR-622 had played with regard to migration and invasion by targeting regulation of DYRK2 pathway in CRC cells. RESULTS: In our research, we found that the expression of miR-622 was elevated in CRC tissues and cell lines compared to that of nonCRC tissues and the normal human colon epithelial cell line NCM460. Correspondingly, the expression of DYRK2 in CRC tissues and cell lines showed a contrary tendency. The different expression level of DYRK2 was closely correlated with clinicopathological characteristics of CRC patients. We demonstrated that down-regulation of miR-622 could inhibit the ability of migration and invasion of CRC cell lines SW1116 and SW480. Also, we confirmed that DYRK2 was negatively regulated by miR-622 via a specific targeted binding site within the 3'UTR. We finally verified that the migration and invasion ability of CRC cells in the conducted DYRK2 3'UTR defect plasmid transfection group were lower compared to miR-622 and cotransfection group. CONCLUSION: The findings of this study indicate that a decrease of miR-622 expression could suppress migration and invasion by targeting regulation of DYRK2 and miR-622/DYRK2 could be a potential molecular treating target of CRC.