DNA Binding Activity Of Transcription Factors Research Articles

The functional and structural characterisation of the bZIP transcription factors from Myristica fragrans Houtt. associated to plant disease-resistant defence: An insight from transcriptomics and computational modelling

The bZIP transcription factors play crucial roles in various aspects of plant biology, including development, defence mechanisms, senescence, and responses to both biotic and abiotic environmental stresses. Myristica fragrans Houtt. transcriptome analysis has identified 15 bZIP transcription factors, each exhibiting major conserved domains and motifs such as BRLZ, MFMR, and DOG1. Functional characterisation of these identified MfbZIP factors indicates their predominant localisation within the nucleus. Phylogenetic analysis reveals that MfbZIP factors cluster into three subgroups alongside annotated bZIP sequences from Magnolia sinica and Arabidopsis thaliana. Moreover, gene ontology (GO) analysis highlights several key functions of MfbZIP, including involvement in defence responses, abscisic acid-induced signalling pathways, and DNA-binding transcription factor activity. Further investigation through KEGG pathway analysis reveals that the amino acid sequences of MfbZIP contain binding motifs for proteins such as TGA, implicated in plant hormone signal transduction pathways associated with disease resistance. To confirm the disease-defence-related activity of the TGA binding protein within MfbZIP, we employed amino acid sequences for 3-D ab initio modelling. Subsequently, we analysed TGA-NPR1 interactions using docking and molecular dynamics simulation analysis. These analyses shed light on the functional and structural aspects of TGA, demonstrating its stable association with NPR1 protein and its significance in the expression of PR1 protein, thus playing a pivotal role in defence responses against pathogens.

Abstract 4892: MicroRNA signature and telomere genes in kidney cancer survival

Abstract Introduction: Kidney cancer ranks among the top 10 most prevalent cancers in the United States, clear cell renal cell carcinoma (ccRCC) has the worst disease specific survival histology. MicroRNAs (miRNAs) have emerged as potential prognostic indicators in various cancers, including kidney cancers. Telomerase, a biomarker associated with uncontrolled cancer cell growth, plays a crucial role in the human kidney cortex's telomere shortening over time. However, the link between miRNAs and telomeres in kidney cancer survival remains underexplored. Our study aims to uncover a miRNA signature capable of predicting 5-year survival in ccRCC patients and to investigate the biological relevance of this signature, particularly its shared pathway enrichment mechanisms with telomere genes. Methods: We extracted miRNA expression profiles from The Cancer Genome Atlas database, comprising 166 patients with ccRCC. To identify a robust miRNA signature predicting 5-year survival in ccRCC, we established an Evolutionary Kidney Survival Estimator (EKSE) using an optimal feature selection algorithm and support vector regression. Additionally, we conducted bioinformatics analyses to uncover common biological pathways between miRNAs and telomere genes using Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) categories, encompassing biological processes, molecular functions, and cellular components. Results: EKSE identified a robust signature consisting of 37 miRNAs, achieving a mean correlation coefficient and mean absolute error of 0.82±0.01 and 0.65±0.18, respectively between actual and estimated survival times. The top 10 ranked miRNAs in the signature, including hsa-miR-26a-1-3p, hsa-miR-28-5p, hsa-miR-3913-5p, hsa-miR-3170, hsa-miR-148a-5p, hsa-miR-671-3p, hsa-miR-224-3p, hsa-miR-10a-5p, hsa-miR-29b-1-5p, and hsa-miR-106b-5p, also showed promise in diagnosing ccRCC with AUC values ranging from 0.56 to 0.91. Furthermore, bioinformatics analysis revealed that the miRNA signature targeted well known telomere-associated genes such as TERT, DKC1, CTC1, and RTEL1. Notably, miRNA signature and telomere genes shared several common pathways, including cellular senescence (hsa04218) and proteoglycans in cancer (hsa05205). They also had overlapping GO-Molecular functions, such as DNA-binding transcription factor activity (GO:0003700), histone deacetylase binding (GO:0042826), and promoter-specific chromatin binding (GO:1990841). Conclusions: Our findings suggest an association between microRNAs and telomere genes, potentially playing significant roles in the survival of patients with ccRCC. The identified miRNA signature robustly predicts 5-year survival in ccRCC. The shared biological pathways between these miRNAs and telomere genes offer new insights into the mechanisms driving ccRCC, signaling potential for therapeutic targets and improved prognostic markers in kidney cancer. Citation Format: Srinivasulu Yerukala Sathipati, Rohit Sharma, Scott Hebbring. MicroRNA signature and telomere genes in kidney cancer survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4892.

Computational Analysis of Single Nucleotide Polymorphisms in Human HIC1 Gene.

Background A putative tumor suppressor gene called HIC1 (hypermethylated in cancer) is situated at 17p13.3, a locus where the allelic loss occurs often in human malignancies, including breast cancer. Hypermethylated in cancer 1 protein is a protein that in humans is encoded by the HIC1 gene and it's aHomo sapiens(Human). This gene functions as a growth regulatory and tumor repressor gene. The molecular function of HIC1 gene includes DNA-binding transcription factor activity, sequence-specific DNA binding, DNA binding, histone deacetylase binding, protein binding, metal ion binding, nucleic acid binding, DNA-binding transcription repressor activity, RNA polymerase II-specific, DNA-binding transcription factor activity, RNA polymerase II-specific. The biological process of HIC1 gene includes multicellular organism development, negative regulation of Wnt signaling pathway, positive regulation of DNA damage response, signal transduction by p53 class mediator regulation of transcription, DNA-templated, negative regulation of transcription by RNA polymerase II, Wnt signaling pathway, transcription, DNA-templated, intrinsic apoptotic signaling pathway in response to DNA damage, cellular response to DNA damage stimulus. The study aimed to predict the stability and structure of the protein that will arise from single nucleotide polymorphisms (SNPs) in the human HIC1 gene. Methodology To investigate the possible negative effects associated with these SNPs, bioinformatic analysis is typically essential. The following tools were employed for forecasting harmful SNPs: scale-invariant feature transform (SIFT), Protein Analysis Through Evolutionary Relationships (PANTHER), nonsynonymous SNP by Protein Variation Effect Analyzer (PROVEAN), and nonsynonymous SNP by Single Nucleotide Polymorphism Annotation Platform (SNAP). Results The present study identified a total of 36 SNPs using the SIFT approach, which were shown to have functional significance. Twenty-six were determined to be tolerable, whereas 10 were shown to be detrimental. Out of 20 SNPs, seven (P370A, P646S, R654P, A476T, S400S, D666N, D7V) SNPs were predicted as "Possibly damaging" and seven (L9F, G468R, G490R, L482R, S12W, G489D, S12P) were identified as "probably benign", and six (R725G, G620S, A56V, E463D, D394N, L338V) were identified as "probably damaging" according to the predictions made by PANTHER tools. The majority of the pixels on the strip were red, indicating that the gene changes may have dangerous consequences. These results highlight the need for more research to fully comprehend how these mutations affect the hic1 protein's function, which is essential for the emergence of different types of cancer. Conclusion The current research has provided us with essential information about how SNPs might be used as a diagnostic marker for cancer, given that SNPs may be candidates for cellular changes caused by mutations linked to cancer.

Expression changes of miRNA-regulated genes associated with the formation of the leafy head in cabbage (Brassica oleracea var. capitata)

The vegetative development of cabbage (B. oleracea var. capitata) passes through seedling, rosette, folding and heading stages. Leaves that form the rosette are large and mostly flat. In the following developmental stages, the plants produce leaves that curve inward to produce the leafy head. Many microRNAs and their target genes have been described participating in leaf development and leaf curvature. The aim of this study is to investigate the role of miRNA-regulated genes in the transition from the rosette to the heading stage. We compared the miRNA and gene abundances between emerging rosette and heading leaves. To remove transcripts (miRNAs and genes) whose regulation was most likely associated with plant age rather than the change from rosette to heading stage, we utilized a non-heading collard green (B. oleracea var. acephala) morphotype as control. This resulted in 33 DEMs and 1 998 DEGs with likely roles in the transition from rosette to heading stage in cabbage. Among these 1 998 DEGs, we found enriched GO terms related to DNA-binding transcription factor activity, transcription regulator activity, iron ion binding, and photosynthesis. We predicted the target genes of these 33 DEMs and focused on the subset that was differentially expressed (1 998 DEGs) between rosette and heading stage leaves to construct miRNA-target gene interaction networks. Our main finding is a role for miR396b-5p targeting two Arabidopsis thaliana orthologues of GROWTH REGULATING FACTORs 3 (GRF3) and -4 (GRF4) in pointed cabbage head formation.

Identification of potential immunotherapeutic targets and prognostic biomarkers in Graves' disease using weighted gene co-expression network analysis

Graves' disease (GD) is an autoimmune disorder characterized by hyperthyroidism resulting from autoantibody-induced stimulation of the thyroid gland. Despite recent advancements in understanding GD's pathogenesis, the molecular processes driving disease progression and treatment response remain poorly understood. In this study, we aimed to identify crucial immunogenic factors associated with GD prognosis and immunotherapeutic response. To achieve this, we implemented a comprehensive screening strategy that combined computational immunogenicity-potential scoring with multi-parametric cluster analysis to assess the immunomodulatory genes in GD-related subtypes involving stromal and immune cells. Utilizing weighted gene co-expression network analysis (WGCNA), we identified co-expressed gene modules linked to cellular senescence and immune infiltration in CD4+ and CD8+ GD samples. Additionally, gene set enrichment analysis enabled the identification of hallmark pathways distinguishing high- and low-immune subtypes. Our WGCNA analysis revealed 21 gene co-expression modules comprising 1,541 genes associated with immune infiltration components in various stages of GD, including T cells, M1 and M2 macrophages, NK cells, and Tregs. These genes primarily participated in T cell proliferation through purinergic signaling pathways, particularly neuroactive ligand-receptor interactions, and DNA binding transcription factor activity. Three genes, namely PRSS1, HCRTR1, and P2RY4, exhibited robustness in GD patients across multiple stages and were involved in immune cell infiltration during the late stage of GD (p < 0.05). Importantly, HCRTR1 and P2RY4 emerged as potential prognostic signatures for predicting overall survival in high-immunocore GD patients (p < 0.05). Overall, our study provides novel insights into the molecular mechanisms driving GD progression and highlights potential key immunogens for further investigation. These findings underscore the significance of immune infiltration-related cellular senescence in GD therapy and present promising targets for the development of new immunotherapeutic strategies.

Breast cancer risk SNPs and associated expression QTLs focusing Bangladeshi population: An in silico analysis

BackgroundSingle Nucleotide Polymorphism (SNP)s in the genome and associated genes cause susceptibility to breast cancer, the most common cancer leading to death in women. Variation in different human races' genomes makes breast cancer prognosis challenging in terms of targeted drugs and therapies. The study aimed to compile the Breast cancer associated SNPs used for screening in the existing publications focusing Bangladeshi population, followed by the identification of Expression quantitative trait loci (eQTLs) associated with those SNPs from the eQTL database. eQTLs identify genes whose expression is regulated by specific SNPs. In silico characterization in terms of variant effect prediction, co-expression, Gene Ontology (GO) enrichment, protein-protein interaction, and sequence motif analysis narrowed down a specific set of candidate genes. MethodsPublished reports emphasizing the SNPs screened for Breast cancer in Bangladeshi population were analyzed in PancanQTL for identification of eQTLs which uses genotype and gene expression data from The Cancer Genome Atlas. The gene description and GO associated with identified eQTLs were retrieved from the Ensembl database and characterizations were performed using variant effect predictor, Coexpedia, MEME suite, and STRINGdb. ResultsIt was found from the published reports that not all variants showed strong association with the disease in Bangladeshi population. The cis-eQTLs associated with reported SNPs tested so far on Bangladeshi population are ZNF575, MRPL42P5, C15orf57, C15orf62, NFATC3, XRCC1, C14orf153, CKB, BAG5, KLC1, MARK3. Among them only ZNF575 was enlisted as breast cancer associated eQTL and the rest are linked with other types of cancer. These genes are mostly associated in DNA-binding transcription factor activity, protein binding, Intracellular protein transport II, transferase activity. Protein-protein interaction could predict some functional partners to connect the eQTLs with respective SNP harboring genes. Taking the commonly screened genes for breast cancer as targets breast cancer associated cis and trans eQTLs along with the associated survival eQTLs have been retrieved from the database and a list of specific variants are recommended for future studies to get a more comprehensive scenario about the disease prognosis. ConclusionSince it was found from the existing literature that the commonly used variants are not always associated with all human races, this simple and precise in silico study was carried out to analyze publicly available data. This helped limit specific candidate genes and variants which will be helpful in future population-based screenings in understanding breast cancer prognosis with a provision to develop population specific personalized drug.

A comprehensive analysis of transcriptomic data for comparison of cold tolerance in two Brassica napus genotypes.

Brassica napus is an important oil crop and cold stress severely limits its productivity. To date, several studies have reported the regulatory genes and pathways involved in cold-stress responses in B. napus. However, transcriptome-scale identification of the regulatory genes is still lacking. In this study, we performed comparative transcriptome analysis of cold-tolerant C18 (CT - C18) and cold-sensitive C6 (CS - C6) Brassica napus genotypes under cold stress for 7 days, with the primary purpose of identifying cold-responsive transcription in B. napus. A total of 6061 TFs belonging to 58 families were annotated in the B. napus genome, of which 3870 were expressed under cold stress in both genotypes. Among these, 451 TFs were differentially expressed (DE), with 21 TF genes expressed in both genotypes. Most TF members of the MYB (26), bHLH (23), and NAC (17) families were significantly expressed in the CT - C18 genotype compared with the CS - C6 B. napus genotype. GO classification showed a significant role in transcription regulation, DNA-binding transcription factor activity, response to chitin, and the ethylene-activated signaling pathway. KEGG pathway annotation revealed these TFs are involved in regulating more pathways, resulting in more tolerance. In conclusion, the results provide insights into the molecular regulation mechanisms of B. napus in response to freezing treatment, expanding our understanding of the complex molecular mechanisms in plants' response to freezing stress.

Beauveria bassiana transcriptomics reveal virulence-associated shifts during insect lipid assimilation.

Insect cuticular lipids, especially epicuticular hydrocarbons (CHC), have a significant role in insect ecology and interactions with other organisms, including fungi. The CHC composition of a specific insect species may influence the outcome of the interaction with a specific fungal strain. Some insects, such as Piezodorus guildinii, have low susceptibility towards fungal infections seemingly due to their CHC composition. The entomopathogenic fungus Beauveria bassiana can assimilate CHC and incorporate them as building blocks via cytochrome P450 monooxygenases (CYPs). However, little is known about other enzymes that promote the degradation/assimilation of these cuticular components. In this study, we performed a transcriptomic analysis to evaluate the in vitro response of two virulence-contrasting B. bassiana strains when grown on three different P. guildinii CHC sources. We found a different expression profile of virulence-related genes, as well as different GO and KEGG parameters enriched at 4 days post-inoculation, which could help account for the intrinsic virulence and for an alkane-priming virulence enhancement effect. The hypovirulent strain predominantly showed higher expression of cuticle penetration genes, including chitinases, proteases, and CYPs, with GO term categories of "heme binding," "monooxygenase activity," and "peroxisome" pathways enriched. The hypervirulent strain showed higher expression of cell wall remodeling and cell cycle genes, and cuticle adhesion and a distinct set of CYPs, with GO categories of "DNA-binding transcription factor activity" and KEGG pathways corresponding to "meiosis-yeast" and "cell cycle" enriched. These results suggest a delay and alternate routes in pathogenicity-related metabolism in the hypovirulent strain in comparison with the hypervirulent strain. KEY POINTS: •Transcriptomics of two B. bassiana strains grown in P. guildinii cuticular components •Virulence-related genes correlated with virulence enhancement towards P. guildinii •Differentially expressed genes, GOs and KEGGs showed different metabolic timelines associated with virulence.

The effect of PK gene overexpression on content and antioxidant properties of carotenoids in marine microalga Dunaliella parva

Dunaliella parva can extensively accumulate carotenoids, which is a promising raw material for carotenoids production. Carotenoids have important medicinal value. D. parva is an ideal organism for studying the mechanism of carotenoid synthesis. Our previous study identified a transcription factor DpAP2 which could regulate carotenoid synthesis in D. parva. In addition, DpAP2 could interact with three proteins with different activities (DNA binding transcription factor activity, protein kinase activity, and alpha-D-phosphohexomutase). To investigate the function of PK gene encoding interacting protein of DpAP2 with protein kinase activity in D. parva, PK gene was cloned into vector pBI221-GFP-UbiΩ-CAT and transformed into D. parva in this study. The results showed that overexpression of PK gene enhanced the contents of carotenoids, total sugars, proteins, and antioxidant activities of carotenoid extract such as superoxide radical scavenging activity, reducing power, hydroxyl radical scavenging activity in transgenic D. parva with overexpression of PK gene. This study explored the function of PK gene, and improved the medicinal value of D. parva.

Transcription factor network analysis of the Cf-19-mediated resistance response in tomato infected by Cladosporium fulvum

Tomato leaf mold, caused by Cladosporium fulvum, is a common disease in tomato cultivation. The Cf-19 gene is an effective resistance gene to C. fulvum. Current studies have shown that the Cf-19-mediated resistance response pathway is regulated by a variety of TFs. In this study, in order to better understand the role of TFs in the Cf-19-mediated immune response of tomato against C. fulvum, differentially expressed TFs in different response stages were identified based on RNA-seq data. The results showed that a total of 37 families with 171 TF members were identified. These differentially expressed TFs were enriched in “nucleus” in the component category, “DNA-binding transcription factor activity” in the function category and “Regulation of transcription” in the process category in the Gene Ontology (GO) analysis results. Four pathways, including plant hormone signal transduction, MAPK signaling, plant-pathogen interaction and circadian rhythm, were enriched in the KEGG analysis results. Further analysis showed that 45 TFs were involved in the above four major pathways, and some TFs in different pathways had interaction relationships, which might be the key nodes mediating cross-talk between different pathways. EREB was predicted to be the core of multigroup interactions, and its interaction with WRKY80 is the node connecting the MAPK pathway and plant-pathogen interaction pathway. The function of the EREB gene was analyzed by VIGS, and it was found that the downregulated expression of this gene decreased the expression of WRKY80 and the disease resistance mediated by Cf-19.

Characterizing Different Modes of Interplay Between Rap1 and H3 Using Inducible H3-depletion Yeast

Histones and transcription factors (TFs) are two important DNA-binding proteins that interact, compete, and together regulate transcriptional processes in response to diverse internal and external stimuli. Condition-specific depletion of histones in Saccharomyces cerevisiae using a galactose-inducible H3 promoter provides a suitable framework for examining transcriptional alteration resulting from reduced nucleosome content. However, the effect on DNA binding activities of TFs is yet to be fully explored. In this work, we combine ChIP-seq of H3 with RNA-seq to elucidate the genome-scale relationships between H3 occupancy patterns and transcriptional dynamics before and after global H3 depletion. ChIP-seq of Rap1 is also conducted in the H3-depletion and control treatments, to investigate the interplay between this master regulator TF and nucleosomal H3, and to explore the impact on diverse transcriptional responses of different groups of target genes and functions. Ultimately, we propose a working model and testable hypotheses regarding the impact of global and local H3 depletion on transcriptional changes. We also demonstrate different potential modes of interaction between Rap1 and H3, which sheds light on the potential multifunctional regulatory capabilities of Rap1 and potentially other pioneer factors.

Evaluating Tartary Buckwheat Genotypes with High Callus Induction Rates and the Transcriptomic Profiling during Callus Formation.

Due to their complex genotypes, low in vitro regeneration rates, and difficulty in obtaining transgenic plants, studies concerning basic biological research and molecular breeding in Tartary buckwheat (TB) are greatly limited. In this study, the hypocotyls of 60 genotypes of TB (TBC1~60) were used as explants. Of these, TBC14 was selected due to a high callus induction rate of 97.78% under dark and a proliferation coefficient (PC) of 28.2 when cultured on MS medium supplemented with 2.0 mg/L of 2,4-D and 1.5 mg/L of 6-BA. Subsequently, the samples of the calli obtained from TBC14 were collected at 0, 10, 20, and 30 d, and their transcriptomes were sequenced where identified. GO enrichment led to the detection of the most significant active gene set, which was the DNA binding transcription factor activity. The DEGs related to the pathways concerning metabolism, the biosynthesis of secondary metabolites, and hormone signal transduction were the most enriched in the KEGG database. The sets of MYB, AP2/ERF, and bHLH TFs exhibited the highest number of DEGs. Using this enrichment analysis, 421 genes encoding TFs, 47 auxin- and cytokinin-related genes, and 6 signal transduction-associated genes were screened that may play significant roles in callus formation (CF) in TB. Furthermore, FtPinG0008123200.01 (bZIP), a key gene promoting CF, was screened in terms of the weighted gene co-expression network associated with the various stages of CF. Our study not only provides valuable information about the molecular mechanism of CF but also reveals new genes involved in this process.

A sticky-end probe biosensor for homogeneous detection of transcription factor binding activity.

Transcription factors are essential regulators of various physiological and pathological processes. However, detecting transcription factor-DNA binding activities is often time-consuming and labor-intensive. Homogeneous biosensors that are compatible with mix-and-measure protocols have the potential to simplify the workflow for therapeutic screening and disease diagnostics. In this study, we apply a combined computational-experimental approach to investigate the design of a sticky-end probe biosensor, where the transcription factor-DNA complex stabilizes the fluorescence resonance energy transfer signal of the donor-acceptor pair. We design a sticky-end biosensor for the SOX9 transcription factor based on the consensus sequence and characterize its sensing performance. A systems biology model is also developed to investigate the reaction kinetics and optimize the operating conditions. Taken together, our study provides a conceptual framework for the design and optimization of sticky-end probe biosensors for homogeneous detection of transcription factor-DNA binding activity.

Deciphering the Role of the Ser-Phosphorylation Pattern on the DNA-Binding Activity of Max Transcription Factor Using Chemical Protein Synthesis.

The chemical synthesis of site-specifically modified transcription factors (TFs) is a powerful method to investigate how post-translational modifications (PTMs) influence TF-DNA interactions and impact gene expression. Among these TFs, Max plays a pivotal role in controlling the expression of 15% of the genome. The activity of Max is regulated by PTMs; Ser-phosphorylation at the N-terminus is considered one of the key regulatory mechanisms. However, the molecular impact of the phosphorylation pattern on the DNA binding and sequence specificities remains elusive. Here, we developed a practical synthetic strategy to prepare homogeneous full-length Max for the first time, to explore the impact of Max phosphorylation. We prepared a focused library of eight Max variants, with distinct modification patterns, including mono-phosphorylated, and doubly-phosphorylated analogs at Ser2/ Ser11 as well as fluorescently-labeled variants via native chemical ligation. Through comprehensive DNA binding analyses, we discovered that the phosphorylation position plays a crucial role in Max's DNA binding activity. Furthermore, in-vitro high-throughput analysis using DNA microarrays revealed that the N-terminus phosphorylation pattern does not interfere with Max's DNA sequence specificity. Our work provides insights into the regulatory role of Max's phosphorylation on the DNA interactions and sequence specificity, shedding light on how PTMs influence TF function.

Rice miR5504 regulates plant height by affecting cell proliferation and expansion.

miRNAs play critical roles in the regulation of plant growth and development by cleaving mRNA or repressing transcription. In our previous study, miR5504 with unknown functions was captured by small RNA sequencing. Here, the function and characters of miR5504 were extensively analyzed using CRISPR/Cas9, overexpression strategy, Northern blot, cytological analysis, and transcriptomics analysis. We found that the dwarf phenotype of mir5504 mutants (mir5504-1 and mir5504-2) appeared on 35-day seedlings and became more apparent at the mature stage. The cytological results showed a substantial decrease in the vascular bundle number, cell number and cell length in the mir5504 mutant compared with NIP. In addition, we found that miR5504 regulated plant height by targeting LOC_Os08g16914. The results of RNA-seq revealed that numerous biological processes were mainly enriched, including DNA-binding transcription factor activity, transferase activity, regulation of transcription, metabolic process, and protein binding. Meanwhile, KEEG analysis showed that numerous proteins were associated with cellular processes and metabolism pathways. Taken together, miR5504 may be involved in the regulation of plant height by affecting cell expansion and division of internode in rice.

Exploring the novel SNPs in neuroticism and birth weight based on GWAS datasets

ObjectivesEpidemiological studies have confirmed that low birth weight (BW) is related to neuroticism and they may have a common genetic mechanism based on phenotypic correlation research. We conducted our study on a European population with 159,208 neuroticism and 289,142 birth weight samples. In this study, we aimed to identify new neuroticism single nucleotide polymorphisms (SNPs) and pleiotropic SNPs associated with neuroticism and BW and to provide more theoretical basis for the pathogenesis of the disease.MethodsWe estimated the pleiotropic enrichment between neuroticism and BW in two independent Genome-wide association studies (GWAS) when the statistical thresholds were Conditional False Discovery Rate (cFDR) < 0.01 and Conjunctional Conditional False Discovery Rate (ccFDR) < 0.05. We performed gene annotation and gene functional analysis on the selected significant SNPs to determine the biological role of gene function and pathogenesis. Two-sample Mendelian Randomization (TSMR) analysis was performed to explore the causal relationship between the neuroticism and BW.ResultsThe conditional quantile–quantile plots (Q-Q plot) indicated that neuroticism and BW have strong genetic pleiotropy enrichment trends. With the threshold of cFDR < 0.001, we identified 126 SNPs related to neuroticism and 172 SNPs related to BW. With the threshold of ccFDR < 0.05, we identified 62 SNPs related to both neuroticism and BW. Among these SNPs, rs8039305 and rs35755513 have eQTL (expressed quantitative trait loci) and meQTL (methylation quantitative trait loci) effects simultaneously. Through GO enrichment analysis we also found that the two pathways of positive regulation of “mesenchymal cell proliferation” and “DNA-binding transcription factor activity” were significantly enriched in neuroticism and BW. Mendelian randomization analysis results indicate that there is no obvious causal relationship between neuroticism and birth weight.ConclusionWe found 126 SNPs related to neuroticism, 172 SNPs related to BW and 62 SNPs associated with both neuroticism and BW, which provided a theoretical basis for their genetic mechanism and novel potential targets for treatment/intervention development.

Identification and Characterization of Development-Related microRNAs in the Red Flour Beetle, Tribolium castaneum

MicroRNAs (miRNAs) play important roles in insect growth and development, but they were poorly studied in insects. In this study, a total of 883 miRNAs were detected from the early embryo (EE), late larva (LL), early pupa (EP), late pupa (LP), and early adult (EA) of Tribolium castaneum by microarray assay. Further analysis identified 179 differentially expressed unique miRNAs (DEmiRNAs) during these developmental stages. Of the DEmiRNAs, 102 DEmiRNAs exhibited stage-specific expression patterns during development, including 53 specifically highly expressed miRNAs and 20 lowly expressed miRNAs in EE, 19 highly expressed miRNAs in LL, 5 weakly expressed miRNAs in EP, and 5 abundantly expressed miRNAs in EA. These miRNAs were predicted to target 747, 265, 472, 234, and 121 genes, respectively. GO enrichment analysis indicates that the targets were enriched by protein phosphorylation, calcium ion binding, sequence-specific DNA binding transcription factor activity, and cytoplasm. An RNA interference-mediated knockdown of the DEmiRNAs tca-miR-6-3p, tca-miR-9a-3p, tca-miR-9d-3p, tca-miR-11-3p, and tca-miR-13a-3p led to defects in metamorphosis and wing development of T. castaneum. This study has completed the identification and characterization of development-related miRNAs in T. castaneum, and will enable us to investigate their roles in the growth and development of insect.

MP41-12 ASSOCIATION OF KIDNEY CANCER WITH BOTH NOVEL AND REPORTED MONOGENIC GENE MUTATIONS: RESULTS OF WHOLE EXOME SEQUENCING IN THE UK BIOBANK

You have accessJournal of UrologyCME1 Apr 2023MP41-12 ASSOCIATION OF KIDNEY CANCER WITH BOTH NOVEL AND REPORTED MONOGENIC GENE MUTATIONS: RESULTS OF WHOLE EXOME SEQUENCING IN THE UK BIOBANK Jun Wei, Kristian Novakovic, Zhuqing Shi, Andrew Rifkin, S. Lilly Zheng, Brian Helfand, and Jianfeng Xu Jun WeiJun Wei More articles by this author , Kristian NovakovicKristian Novakovic More articles by this author , Zhuqing ShiZhuqing Shi More articles by this author , Andrew RifkinAndrew Rifkin More articles by this author , S. Lilly ZhengS. Lilly Zheng More articles by this author , Brian HelfandBrian Helfand More articles by this author , and Jianfeng XuJianfeng Xu More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003279.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Pathogenic mutations in several cancer susceptibility genes have been reported in kidney cancer patients. However, their associations with kidney cancer have not been formally tested in well-characterized case-control studies and using the same sequencing and analytical pipeline. The objective of this study was to systematically evaluate association of kidney cancer with genes in the exome in a well-characterized prospective population-based case-control cohort. METHODS: A total of 187,966 subjects from the UK Biobank (UKB) with whole exome sequencing data were included in the analysis, including 1,217 kidney cancer patients (ICD10 C64). Pathogenic/likely pathogenic (P/LP) mutations were annotated based on criteria of the American College of Medical Genetics. The robust SKAT-O, a novel gene-based analysis that properly controls for type I error rates due to unbalanced case-control ratio, was used for association tests adjusting for age at recruitment, gender and genetic background. RESULTS: Among the 21 previously reported candidate genes, three were statistically significant at P<0.05, including VHL, SDHB, and CHEK2 (Table). In addition, associations with 9 novel genes (at P<0.0001) were found, including ZNF772, PRIMPOL, HES4, MOXD1, KRTAP10-9, ZNF827, ACAN, CDK20, and LRRFIP1. While these associations were only marginally significant after adjusting for multiple testing, they have potential roles in cancer development. For example, ZNF772 is predicted to enable DNA-binding transcription factor activity, RNA polymerase II-specific and RNA polymerase II cis-regulatory region sequence-specific DNA binding activity which are all known to be dysregulated in cancer. CONCLUSIONS: Identification of genes that influence kidney cancer may advance our understanding of disease etiology and identify potential therapeutic targets. Using the same sequencing method and mutation annotation pipeline in the largest population-based cohort to date, we confirmed several previously reported genes and identified several novel candidate genes for their association with kidney cancer. Confirmation in independent study populations is required for these novel candidate genes. Source of Funding: None. © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e559 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jun Wei More articles by this author Kristian Novakovic More articles by this author Zhuqing Shi More articles by this author Andrew Rifkin More articles by this author S. Lilly Zheng More articles by this author Brian Helfand More articles by this author Jianfeng Xu More articles by this author Expand All Advertisement PDF downloadLoading ...

Time-course RNA-seq analysis provides an improved understanding of genetic regulation in response to cold stress from white clover (Trifolium repens L.)

White clover (Trifolium repens L.) is an important legume forage, which is widely distributed in cool-season regions. Therefore, cold stress is a major environmental factor limiting its growth and production, while little is known about the its cold tolerance at molecular level. In the present study, we performed time-course RNA-seq analysis under cold stress. RNA-seq results suggested that genes associated with “oxidoreductase activity” and “transcription regulator activity” are more likely related to the response of white clover to cold stress. To identify the specific gene modules and the hub genes of white clover in response to cold stress, we applied weighted gene co-expression network analyses (WGCNA) to transcriptome data. We also found that gene modules that focus on protein kinase activity, DNA-binding transcription factor activity and oxidoreductase activity, are more likely to be involved in the response of white clover to cold stress. Especially, we identified several AP2/ERF TF genes and CDPK genes as pivotal genes in white clover in response to cold stress, which would provide helpful insights into the molecular mechanisms underlying the response of white clover to cold stress.

Transcriptomic analysis reveals the mechanism of the alleviation of salt stress by salicylic acid in pepper (Capsicum annuum L.)

BackgroundThe growth and yield of pepper (Capsicum annuum L.) is often affected by the critical salt stress. Salicylic acid (SA) can improve plants’ stress tolerance by promoting growth and regulating ion absorption and transportation.Methods and resultsTo uncover the alleviated mechanism of salt stress by SA in pepper, we conducted morphological, physiological, cytological, and transcriptomic analyses under a single SA treatment and NaCl with and without SA pre-treatment for 9 days. Seedlings under NaCl treatment showed yellow shrunken leaves, this tatus were alleviated by NS treatment (NaCl with SA pre-treatment). Compared with plants under NaCl treatment, those in the NS treatment showed reduced lipid peroxidation, and significantly increased contents of chlorophyll and osmotic regulators (proline, soluble sugars). Treatment with SA balanced the Na+/K+ ratio. We conducted transcriptome sequencing and identified differentially expressed genes (DEGs) contributing to alleviation of salt stress by SA in pepper. Besides photosynthesis related genes, GO and KEGG analyses revealed that the DEGs were enriched in ‘sequence-specific DNA binding’, ‘transcription regulator activity’ and ‘DNA binding transcription factor activity’ by GO terms. And our results showed that TFs, such as MYB, bZIP, BBX, AP2/ERF, NAC, etc., probably make a great contribution in the alleviation of salt stress by SA.ConclusionsThese results reveal that SA can improve plants’ stress tolerance by balancing ion absorption, gene expression and transcriptional regulation, which provide new ideas and resources for subsequent research on the mechanism of salt tolerance in pepper.